HQ-O Ready-To-Dilute (RTD TM ) Stain Reagent is designed to label amyloid plaques in paraffin-embedded or freshly cut frozen tissue sections. As a fluorescent zinc chelator, HQ-O is unique as it takes advantage of the known presence of concentrated zinc in amyloid plaques. Studies with HQ-O revealed that fluorescent plaque-like structures are only seen when synthetic A?x-42 is aggregated in the presence of zinc. Under blue light excitation, plaque structures appear bright green fluorescent in the brain parenchyma, correlating closely with plaque structures observed following A? antibody staining. HQ-O RTD TM staining reagent is compatible with other fluorophores, such as DAPI, Hoechst and ethidium bromide, as well as fluorescent-labelled antibodies with emission spectra in the blue and/or red emission range of fluorescent microscopes. Due to its zinc-chelating characteristics, HQ-O RTD TM staining reagent may visualize globular structures within blood vessels and intravascular leucocytes. HQ-O RTD TM staining reagent has multiple advantages over older blue-light exciting stains such as Thioflavin S. Thioflavin S typically exhibits relatively low contrast and resolution and suffers from bleed-through when excited by wavelengths other than blue light. HQ-O RTD TM staining reagent suffers none of these setbacks and not only provides a higher contrast and longer lasting dye, but because it lacks excitation bleed-through, HQ-O can be readily adapted to multiple labelling studies very easily. To visualize the HQ-O tracer, it is recommended to use a filter cube designed for visualizing Fluorescein/FITC or a blue-light laser. Although it can be seen with both narrow and wide-band pass filters, there is no need to use a narrow band filter since the compound does not bleed through when excited with other filters. A recommended excitation range of a wide band filter is 447-503 nm, with a peak at 475.
Background Info:
A novel zinc chelator, HQ-O, was developed for localizing zinc within amyloid plaques. The histology involves incubating tissue sections in a dilute aqueous solution of HQ-O.
Detection and fluorescent-staining of amyloid plaques in paraffin-embedded or freshly cut frozen tissue sections, please see detailed protocol for specific use instructions.
Alternative Names:
HQ-O tracer
Kit Components:
One bottle containing 20 mL of 10X HQ-O RTD TM solution This quantity will be sufficient for approximately 4 Coplin Jars or 1-2 staining dishes.
Reconstitution:
Ready to Dilute
Storage:
Store 10X stock solution at 2-8°C protected from light, for up to 6 months. The diluted dye (1X) should be used within 24 hours.
Shelf Life:
6 months after date of receipt (10X stock solution)
Specificity:
As a fluorescent zinc chelator, HQ-O is unique as it takes advantage of the known presence of concentrated zinc in amyloid plaques. Studies with HQ-O revealed that fluorescent plaque-like structures are only seen when synthetic A?x-42 is aggregated in the presence of zinc. Under blue light excitation, plaque structures appear bright green fluorescent in the brain parenchyma, correlating closely with plaque structures observed following A? antibody staining.
Excitation Emission:
To visualize the HQ-O tracer, it is recommended to use a filter cube designed for visualizing Fluorescein/FITC or a blue-light laser. Although it can be seen with both narrow and wide-band pass filters, there is no need to use a narrow band filter since the compound does not bleed through when excited with other filters. A recommended excitation range of a wide band filter is 447 503 nm, with a peak at 475 nm.
HQ-O Ready-To-Dilute (RTD TM ) Stain Reagent is designed to label amyloid plaques in paraffin-embedded or freshly cut frozen tissue sections. As a fluorescent zinc chelator, HQ-O is unique as it takes advantage of the known presence of concentrated zinc in amyloid plaques. Studies with HQ-O revealed that fluorescent plaque-like structures are only seen when synthetic A?x-42 is aggregated in the presence of zinc. Under blue light excitation, plaque structures appear bright green fluorescent in the brain parenchyma, correlating closely with plaque structures observed following A? antibody staining. HQ-O RTD TM staining reagent is compatible with other fluorophores, such as DAPI, Hoechst and ethidium bromide, as well as fluorescent-labelled antibodies with emission spectra in the blue and/or red emission range of fluorescent microscopes. Due to its zinc-chelating characteristics, HQ-O RTD TM staining reagent may visualize globular structures within blood vessels and intravascular leucocytes. HQ-O RTD TM staining reagent has multiple advantages over older blue-light exciting stains such as Thioflavin S. Thioflavin S typically exhibits relatively low contrast and resolution and suffers from bleed-through when excited by wavelengths other than blue light. HQ-O RTD TM staining reagent suffers none of these setbacks and not only provides a higher contrast and longer lasting dye, but because it lacks excitation bleed-through, HQ-O can be readily adapted to multiple labelling studies very easily. To visualize the HQ-O tracer, it is recommended to use a filter cube designed for visualizing Fluorescein/FITC or a blue-light laser. Although it can be seen with both narrow and wide-band pass filters, there is no need to use a narrow band filter since the compound does not bleed through when excited with other filters. A recommended excitation range of a wide band filter is 447-503 nm, with a peak at 475.
Background Info:
A novel zinc chelator, HQ-O, was developed for localizing zinc within amyloid plaques. The histology involves incubating tissue sections in a dilute aqueous solution of HQ-O.
Detection and fluorescent-staining of amyloid plaques in paraffin-embedded or freshly cut frozen tissue sections, please see detailed protocol for specific use instructions.
Alternative Names:
HQ-O tracer
Kit Components:
One bottle containing 40 mL of 10X HQ-O RTD TM solution This quantity will be sufficient for approximately 8 Coplin Jars or 2-4 staining dishes.
Reconstitution:
Ready to Dilute
Storage:
Store 10X stock solution at 2-8°C protected from light, for up to 6 months. The diluted dye (1X) should be used within 24 hours.
Shelf Life:
6 months after date of receipt (10X stock solution)
Specificity:
As a fluorescent zinc chelator, HQ-O is unique as it takes advantage of the known presence of concentrated zinc in amyloid plaques. Studies with HQ-O revealed that fluorescent plaque-like structures are only seen when synthetic A?x-42 is aggregated in the presence of zinc. Under blue light excitation, plaque structures appear bright green fluorescent in the brain parenchyma, correlating closely with plaque structures observed following A? antibody staining.
Excitation Emission:
To visualize the HQ-O tracer, it is recommended to use a filter cube designed for visualizing Fluorescein/FITC or a blue-light laser. Although it can be seen with both narrow and wide-band pass filters, there is no need to use a narrow band filter since the compound does not bleed through when excited with other filters. A recommended excitation range of a wide band filter is 447 503 nm, with a peak at 475 nm.
The Biosensis AG-400-AG kit utilizes an ethidium bromide counter stain for a quick and effective way to visualize cell nuclei and cell bodies of cells while under UV illumination allowing the assessment of amyloid plaques and cell/tissue positioning as well in one step. Amylo-Glo RTD Ready to Dilute Staining reagent is designed to stain amyloid plaques in tissue sections. This novel marker has several advantages over other conventional markers such as Thioflavin S and Congo Red because of its unique chemical and spectral properties. (L. Schmued et al. (2012) J.Neuroscience Methods 209:120- 126). Using Amylo-Glo results in a very bright blue UV excitable stain under physiological conditions that will not bleed through when illuminated with other filters. Its brightness makes it ideal for low magnification quantification studies, while its unique excitation/emission profile and mild staining conditions makes it ideal for combination for multiple immunofluorescent labeling studies. Amylo-Glo RTD is compatible with fresh, frozen, and formalin-fixed immunohistochemistry or cytochemistry, and it is particularly good for confocal and multiple labeling because of its high fluorescent intensity and high resistance to photo-bleaching. Moreover because Amylo-Glo fluoresces in the UV channel, double and triple labeling experiments can be performed very easily (see protocol).
Product Type:
Staining Reagent
Format:
The reagents in the Amyloid Plaque Stain Reagent (100x) are all supplied in a liquid format and are ready-to-dilute.
Species Reactivity:
Human,Mouse,Other Mammals (Predicted),Rat
Applications:
ICC,IHC-Frozen,IHC-Paraffin-embedded
Application Details:
Staining of amyloid plaques in human and animal tissues, see included protocol. EtBr counter stain stains nuclei and cell bodies for easy identification and spacial orientation.
Alternative Names:
AmyloGlo
Kit Components:
1 bottle containing 40 mL of 10X Amylo-Glo RTD (A-G RTD) solution 1 bottle containing 40 mL of 10X A-G RTD Ethidium Bromide (EtBr RTD) solution
Reconstitution:
Ready to dilute per protocol 10X solutions
Storage:
The stock solution can be stored for up to 6 months at 2-8°C protected from light. No preservatives. Use sterile technique when handling and proper laboratory procedures.
Shelf Life:
6 months after date of receipt (unopened vial).
Specificity:
Amyloid plaques both intraneuronal and vascular for A-G, Etbr, nuclei and cell bodies both DNA and RNA label
Excitation Emission:
Excitation Peak: 334 nm; Emission Peak: 533 nm - unbound, 438 nm when bound to amyloid. To visualize Amylo-glo in tissue, UV light is required. For example, Amylo-Glo tissue can be examined using an epifluoresent microscope with UV (Nikon UV-2A) filter cube. Excitation (325-375 nm) Emission (400-450 nm) is typical. Also note, it is not uncommon for Amylo-Glo to appear light yellow when examined by eye, yet appear a light blue color when photographed. Visualization of EtBr: Ethidium bromide has an excitation peak of 300 nm and an emission peak 595 nm. Most UV compatible filter sets can be used.
Emre C et al. (2020) "Receptors for pro-resolving mediators are increased in Alzheimer's disease brain." Brain Pathol. [Epub ahead of print]; Application: IHC/IF Species: HumanHascup KN et al. (2019) "LY379268 Does Not Have Long-Term Procognitive Effects nor Attenuate Glutamatergic Signaling in A?PP/PS1 Mice." J Alzheimers Dis. [Epub ahead of print]; Application: IHC/IF Species: MouseHascup ER et al. (2018) "Diet-Induced Insulin Resistance Elevates Hippocampal Glutamate as well as VGLUT1 and GFAP Expression in A?PP/PS1 Mice." J Neurochem. [Epub ahead of print]; Application: IHC/IF Species: Mouse
Research Areas:
Biosensis Stains & Tracing Reagents Amyloid beta research
Related Products:
TR-300,M-1586
Purity:
Thin layer chromatography using alumina plates and a solvent system of ethanol and water (3:1) revealed the presence of two fluorescent isomers. No amount of starting material was detected.
Amylo-Glo RTD Ready to Dilute Staining reagent is designed to stain amyloid plaques in tissue sections. This novel marker has several advantages over other conventional markers such as Thioflavin S and Congo Red because of its unique chemical and spectral properties. (L. Schmued et al. (2012) J.Neuroscience Methods 209:120- 126). Using Amylo-Glo results in a very bright blue UV excitable stain under physiological conditions that will not bleed through when illuminated with other filters. Its brightness makes it ideal for low magnification quantification studies, while its unique excitation/emission profile and mild staining conditions makes it ideal for combination for multiple immunofluorescent labeling studies. Amylo-Glo RTD is compatible with fresh, frozen, and formalin-fixed immunohistochemistry or cytochemistry, and it is particularly good for confocal and multiple labeling because of its high fluorescent intensity and high resistance to photo-bleaching. Moreover because Amylo-Glo fluoresces in the UV channel, double and triple labeling experiments can be performed very easily (see protocol).
Product Type:
Staining Reagent
Format:
The reagents in the Amyloid Plaque Stain Reagent (100x) are all supplied in a liquid format and are ready-to-dilute.
Species Reactivity:
Human,Mouse,Other Mammals (Predicted),Rat
Applications:
ICC,IHC-Frozen,IHC-Paraffin-embedded
Application Details:
Staining of amyloid plaques in human and animal tissues, see included protocol
Alternative Names:
AmyloGlo
Kit Components:
5 mL of 100X Amylo-Glo RTD (A-G RTD) solution
Reconstitution:
Ready to dilute per protocol; 100X
Storage:
The stock solution can be stored for up to 6 months after date of receipt at 2-8°C protected from light. No preservatives. Use sterile technique when handling and proper laboratory procedures.
Shelf Life:
6 months after date of receipt (unopened vial).
Specificity:
Amyloid plaques both intraneuronal and vascular
Excitation Emission:
Excitation Peak: 334 nm; Emission Peak: 533 nm - unbound, 438 nm when bound to amyloid. To visualize Amylo-glo in tissue, UV light is required. For example, Amylo-Glo tissue can be examined using an epifluoresent microscope with UV (Nikon UV-2A) filter cube. Excitation (325-375 nm) Emission (400-450 nm) is typical. Also note, it is not uncommon for Amylo-Glo to appear light yellow when examined by eye, yet appear a light blue color when photographed.
Da Mesquita S et al. (2021) "Meningeal lymphatics affect microglia responses and anti-A? immunotherapy." Nature. 593(7858):255-260; Application: IHC/IF Species: MouseLauterborn JC et al. (2021) "Increased excitatory to inhibitory synaptic ratio in parietal cortex samples from individuals with Alzheimer's disease. Nat Commun. 12(1):2603; Application: IHC/IF Species: HumanKim JH et al. (2021) "Gamma subunit of complement component 8 is a neuroinflammation inhibitor ." Brain. 144(2):528-552; Application: IHC/IF Species: MouseClaes C et al. (2021) "Plaque-associated human microglia accumulate lipid droplets in a chimeric model of Alzheimer's disease." Mol Neurodegener. 16(1):50; Application: IHC/IF Species: MouseCrapser JD. (2021) "Investigating microglial regulation of the extracellular matrix in health and neurodegenerative disease." PhD Thesis ; Application: IHC/IF Species: HumanBaglietto-Vargas D et al. (2021) "Generation of a humanized A? expressing mouse demonstrating aspects of Alzheimer's disease-like pathology." Nature Communications. 2(1):2421; Application: IHC/IF Species: MouseMistrik M et al. (2021) "Microthermal-induced subcellular-targeted protein damage in cells on plasmonic nanosilver-modified surfaces evokes a two-phase HSP-p97/VCP response." Nature Communications. 12, Article Number 719; Application: ICC/IF Species: HumanLemoine L et al. (2020) "Regional binding of tau and amyloid PET tracers in Down syndrome autopsy brain tissue." Mol Neurodegener. 15(1):68; Application: IHC/IF Species: HumanHascup KN et al. (2020) "Riluzole attenuates glutamatergic tone and cognitive decline in A?PP/PS1 mice." J Neurochem. [Epub ahead of print]; Application: IHC/IF Species: MouseHolloway OG et al. (2020) "Microglia Demonstrate Local Mixed Inflammation and a Defined Morphological Shift in an APP/PS1 Mouse Model. J Alzheimers Dis. 77(4):1765-81; Application: IHC/IF Species: MouseMcQuade A et al. (2020) "Gene expression and functional deficits underlie TREM2-knockout microglia responses in human models of Alzheimer s disease. Nat Commun. 11(1):5370; Application: IHC/IF Species: MouseHascup KN et al. (2020) "Hippocampal alterations in glutamatergic signaling during amyloid progression in A?PP/PS1 mice." Sci Rep. 10(1):14503; Application: IHC/IF Species: MouseCrapser JD et al. (2020) "Microglia facilitate loss of perineuronal nets in the Alzheimer's disease brain." EBioMedicine. 58:102919; Application: IHC/IF Species: MouseAbe Y et al. (2020) "Behavioral and electrophysiological evidence for a neuroprotective role of aquaporin-4 in the 5xFAD transgenic mice model." Acta Neuropathol Commun. 8(1):67; Application: IHC/IF Species: MouseZhu X et al. (2020) "Robust neuroinflammation and perivascular pathology in rTg-DI rats, a novel model of microvascular cerebral amyloid angiopathy." J Neuroinflammation. 17(1):78; Application: IHC/IF Species: RatMajewski L et al. (2020) "Transgenic Mice Overexpressing Human STIM2 and ORAI1 in Neurons Exhibit Changes in Behavior and Calcium Homeostasis but Show No Signs of Neurodegeneration." Int J Mol Sci. 21(3); Application: IHC/IF Species: MouseDavtyan H et al. (2019) "Testing a MultiTEP-based combination vaccine to reduce A? and tau pathology in Tau22/5xFAD bigenic mice."Alzheimers Res Ther. 11(1):107; Application: IHC/IF Species: MouseYeh SHH et al. (2019) "A high-sucrose diet aggravates Alzheimer's disease pathology, attenuates hypothalamic leptin signaling, and impairs food-anticipatory activity in APPswe/PS1dE9 mice."Neurbiol. Aging. [In press]; Application: IHC/IF Species: MouseBharani KL et al. (2019) "Serum Pro-Bdnf Levels Correlate With Phospho-Tau Staining In Alzheimer's Disease."Neurbiol. Aging. [In press]; Application: IHC/IF Species: HumanHovakimyan A et al. (2019) "A MultiTEP platform-based epitope vaccine targeting the phosphatase activating domain (PAD) of tau: therapeutic efficacy in PS19 mice."Sci Rep. 9(1):15455; Application: IHC/IF Species: HumanHasselmann J et al. (2019) "Development of a Chimeric Model to Study and Manipulate Human Microglia In Vivo."Neuron. [Epub ahead of print]; Application: IHC/IF Species: MouseSpangenberg E et al. (2019) "Sustained microglial depletion with CSF1R inhibitor impairs parenchymal plaque development in an Alzheimer's disease model."Nat Commun. 10(1):3758 (Supplementary Figure 1); Application: IHC/IF Species: HumanEggers C et al. (2019) "Novel cannabis flavonoid, cannflavin A displays both a hormetic and neuroprotective profile against amyloid _-mediated neurotoxicity in PC12 cells: comparison with geranylated flavonoids, mimulone and diplacone."Biochem Pharmacol. [Epub ahead of print]; Application: IHC/IF Species: RatDominguez E (2019) "Microglial Contributions to Alzheimer's Disease Pathogenesis." PhD Thesis, UC Irvine. Application: IHC/IF Species: MouseJovic M et al. (2019) "Short-term fish oil supplementation applied in presymptomatic stage of Alzheimer's disease enhances microglial/macrophage barrier and prevents neuritic dystrophy in parietal cortex of 5xFAD mouse model."PLoS One. 14(5):e0216726; Application: IHC/IF Species: MouseCollins MJ et al. (2019) "Age moderates the effects of traumatic brain injury on beta-amyloid plaque load in APP/PS1 mice."J Neurotrauma. [Epub ahead of print]; Application: IHC/IF Species: MouseShukla AK et al. (2018) "CD11a expression distinguishes infiltrating myeloid cells from plaque-associated microglia in Alzheimer's disease."Glia. [Epub ahead of print]; Application: IHC/IF Species: MouseFeng X et al. (2018) "Quantitative proteomics reveals distinct composition of amyloid plaques in Alzheimer's disease."Alzheimers Dement. [In press]; Application: IHC/IF Species: Human, mouseDavis J et al. (2018) "A Novel Transgenic Rat Model of Robust Cerebral Microvascular Amyloid with Prominent Vasculopathy."Am J Pathol. [Epub ahead of print]; Application: IHC/IF Species: RatPalombo F et al. (2017) "Detection of A? plaque-associated astrogliosis in Alzheimer's disease brain by spectroscopic imaging and immunohistochemistry."Analyst. [Epub ahead of print]; Application: IF Species: MouseAbud EM (2017) "Generation of Human Microglia from Induced Pluripotent Stem Cells to Study Innate Immunity in Neurological Diseases."PhD Thesis. 2017; Application: IF Species: MouseAbud EM et al. (2017) "iPSC-Derived Human Microglia-like Cells to Study Neurological Diseases."Neuron. 2017; 49(2):278-93 Application: IF Species: MouseSolomon IH et al. (2017) "Brain and liver pathology, amyloid deposition, and interferon responses among older HIV-positive patients in the late HAART era."BMC Infect Dis. 2017; 17(1):151 Application: IF Species: HumanXu F et al. (2016) "Cerebral vascular amyloid seeds drive amyloid _-protein fibril assembly with a distinct anti-parallel structure."Nat Commun. 2016; 7:13527. Application: IF Species: MouseKatsouri L et al. (2016) "PPARγ-coactivator-1_ gene transfer reduces neuronal loss and amyloid-_ generation by reducing _-secretase in an Alzheimer's disease model ."Proc Natl Acad Sci USA. 2016; 113(43):12292-97. Application: IF Species: MouseEsposito G et al. (2016) "Autologous transplantation of intestine-isolated glia cells improves neuropathology and restores cognitive deficits in _ amyloid-induced neurodegeneration."Sci Rep. 2016; 6: 22605. Application: IF Species: RatMarsh SE et al. (2016) "The adaptive immune system restrains Alzheimer's disease pathogenesis by modulating microglial function."Proc Natl Sci USA. Feb 16. pii: 201525466. Application: IF Species: Hu Fibrillar amyloid visualization.Kim YH et al. (2015) "A 3D human neural cell culture system for modeling Alzheimer's disease."Nat Protoc. Jul;10(7):985-1006. Application: IF Species: Hu, Human neural stem-cell-derived three-dimensional (3D) culture system.Nijholt DA et al. (2015) "Pregnancy Zone Protein is Increased in the Alzheimer's Disease Brain and Associates with Senile Plaques."J Alzheimer's Disease. 46(1):227-38. Application: IF Species: HuKamphuis W et al. (2015) "GFAP and vimentin deficiency alters gene expression in astrocytes and microglia in wild-type mice and changes the transcriptional response of reactive glia in mouse model for Alzheimer's disease."Glia. Jun;63(6):1036-56. Application: IF Species: MouseChoi SH et al. (2014) "A three-dimensional human neural cell culture model of Alzheimer's disease."Nature Oct 12. doi: 10.1038/nature1380. Application: IF Species: Hu, Human neural stem-cell-derived three-dimensional (3D) culture system.Niedowicz DM et al. (2014). "Obesity and diabetes cause cognitive dysfunction in the absence of accelerated beta-amyloid deposition in a novel murine model of mixed or vascular dementia." Acta Neuropathol Commun. 2014 Jun 10;2:64.
Research Areas:
Biosensis Stains & Tracing Reagents Amyloid beta research
Related Products:
TR-400,M-1586
Purity:
Thin layer chromatography using alumina plates and a solvent system of ethanol and water (3:1) revealed the presence of two fluorescent isomers. No amount of starting material was detected.
The causes and effects of neuronal degeneration are of major interest to a wide variety of neuroscientists. Paralleling this growing interest is an increasing number of methods applicable to the detection of neuronal degeneration. Fluoro-Jade C stains all degenerating neurons regardless of specific insult or mechanism of cell death. Fluoro-Jade C exhibits the greatest signal to background ratio, as well as the highest resolution. This translates to a stain of maximal contrast and affinity for degenerating neurons. This makes it ideal for localising not only degenerating nerve cell bodies but also distal dendrites, axons and terminals. The dye is highly resistant to fading and is compatible with virtually all histological processing and staining protocols. Note: This product is equivalent to discontinued product AG325 from Merck-Millipore.
Product Type:
Staining Reagent
Format:
Dry, Coffee brown to brick red powder; hygroscopic powder keep dessicated.
Species Reactivity:
Human,Mouse,Other Mammals (Predicted),Rat
Applications:
FC,ICC
Application Details:
Following our detailed protocol, Fluoro-Jade B labels degenerating neurons which are visualised with blue light excitation, while DAPI (not included) counter stains cell nuclei, visualised with ultra-violet illumination. The Fluoro-Jade C dye can be used on all kinds of preserved tissues, including fresh-frozen, paraformaldehyde or formalin fixed, and formalin fixed, paraffin-embedded tissues.
Alternative Names:
FJC, Fluoro-Jade
Kit Components:
Materials Provided: 30 mg Fluoro-Jade C, dry powder Detailed protocol Equipment and Reagents Required: Distilled water ACS grade Ethanol (200 proof) for slide & solution preparation 1% sodium hydroxide in 80% ethanol (basic alcohol solution) 0.1% Acetic Acid solution (in water) 70% ethanol in distilled water 0.06% (KMnO4) potassium permanganate solution DAPI powder or 100X solution (working range is 0.5-5 µg/mL) Xylene liquid Staining dishes/Coplin jars Cover slips DPX mounting media or another permanent mounting medium. Non-polar media are preferred over aqueous mounting media such as glycerin/water to obtain high- contrast images (refer to Appendix B in the protocol for a comparative analysis). Traditional fluorescent mounting mediums are not recommended because of their high pH. Slide warmer Convection oven
Reconstitution:
Spin vial briefly before opening. See product protocol for detailed use instructions.
Storage:
The powdered dye can be stored desiccated at room temperature in the dark. Storage in a desiccator is recommended as FJB is hydroscopic. The 0.01% stock solution will remain stable for 3 months when stored in a refrigerator, in the dark. The 0.0001-0.0004% working solution in 0.1% acetic acid should be used within 4 hours of preparation. Diluted FJB dye solutions are not stable and should not be stored. The other diluted solutions can be reused and stored for up to 48 hours if refrigerated and protected from light. Best results require freshly diluted solutions.
Shelf Life:
6 months after date of receipt (unopened vial).
Specificity:
Degenerating neurons, and neuronal degeneration. There is no specific staining in normal healthy brain. Note: Some researchers under some conditions report blood vessel staining with Fluoro Jade. This may be because Fluoro Jade is an analogue of eosin (which stains blood cells). In general, good perfusion and preparation of the tissue should help prevent blood vessel staining but it may not be possible to eliminate it entirely. In our experience it is generally possible to distinguish neuronal from blood vessels staining by eye.
Excitation Emission:
FJC visualization is accomplished using blue light or a 488 nm Laser. Excitation Peak: 495 nm Emission Peak: 521 nm Filter system for visualizing: Fluorescein/FITC
TR-100,TR-150 This product TR-160-FJC is equivalent to discontinued product AG325 (FJC) from Merck-Millipore also see "Ready to dilute" RTD FJC : TR-100-FJT and TR-100-FJ
Purity:
Silica TLC (acetanitrile/water, 6/4) revealed the presence of 2 fluorescent spots, presumably corresponding to the mono and di sulphate homologues. The presence of precursors or free fluorescein was not detected.
The causes and effects of neuronal degeneration are of major interest to a wide variety of neuroscientists. Paralleling this growing interest is an increasing number of methods applicable to the detection of neuronal degeneration. The fluorescent dye Fluoro-Jade B (FJB), like its more purified brother Fluoro-Jade C (FJC), is an anionic fluorescein derivative useful for the histological staining of neurons undergoing degeneration. Fluoro-Jade B differs from FJC in that it is a slightly less refined chemical formulation and thus it does not quite provide the same level of signal to noise or high resolution as FJC. Nonetheless FJB is still widely used and works very well as a marker of degenerating neurons and even glia (see Damjanac M et al., Brain Res. 2007;1128(1):40-9). FJB operates nearly identically in protocol to that of FJC, and Fluoro-Jade B is compatible with several other labeling procedures including immunofluorescent and fluorescent Nissl techniques. Fluoro-Jade B stains all degenerating neurons regardless of specific insult or mechanism of cell death. Fluoro-Jade B exhibits the greatest signal to background ratio, as well as the highest resolution. This translates to a stain of maximal contrast and affinity for degenerating neurons. This makes it ideal for localising not only degenerating nerve cell bodies but also distal dendrites, axons and terminals. The dye is highly resistant to fading and is compatible with virtually all histological processing and staining protocols. Note: This product is equivalent to discontinued product AG310 from Merck-Millipore.
Product Type:
Staining Reagent
Format:
Dry, Coffee brown to brick red powder; hygroscopic powder keep dessicated.
Species Reactivity:
Human,Mouse,Other Mammals (Predicted),Rat
Applications:
FC,ICC
Application Details:
Following our detailed protocol, Fluoro-Jade C labels degenerating neurons which are visualised with blue light excitation, while DAPI (not included) counter stains cell nuclei, visualised with ultra-violet illumination. The Fluoro-Jade C dye can be used on all kinds of preserved tissues, including fresh-frozen, paraformaldehyde or formalin fixed, and formalin fixed, paraffin-embedded tissues.
Alternative Names:
FJB, Fluoro-Jade
Kit Components:
Materials Provided: 30 mg Fluoro-Jade B, dry powder Detailed protocol Equipment and Reagents Required: Distilled water ACS grade Ethanol (200 proof) for slide & solution preparation 1% sodium hydroxide in 80% ethanol (basic alcohol solution) 0.1% Acetic Acid solution (in water) 70% ethanol in distilled water 0.06% (KMnO4) potassium permanganate solution DAPI powder or 100X solution (working range is 0.5-5 µg/mL) Xylene liquid Staining dishes/Coplin jars Cover slips DPX mounting media or another permanent mounting medium. Non-polar media are preferred over aqueous mounting media such as glycerin/water to obtain high- contrast images (refer to Appendix B in the protocol for a comparative analysis). Traditional fluorescent mounting mediums are not recommended because of their high pH. Slide warmer Convection oven
Reconstitution:
Spin vial briefly before opening. See product protocol for detailed use instructions.
Storage:
The powdered dye can be stored desiccated at room temperature in the dark. Storage in a desiccator is recommended as FJB is hydroscopic. The 0.01% stock solution will remain stable for 3 months when stored in a refrigerator, in the dark. The 0.0001-0.0004% working solution in 0.1% acetic acid should be used within 4 hours of preparation. Diluted FJB dye solutions are not stable and should not be stored. The other diluted solutions can be reused and stored for up to 48 hours if refrigerated and protected from light. Best results require freshly diluted solutions.
Shelf Life:
6 months after date of receipt (unopened vial).
Specificity:
Degenerating neurons, and neuronal degeneration. There is no specific staining in normal healthy brain. Note: Some researchers under some conditions report blood vessel staining with Fluoro Jade. This may be because Fluoro Jade is an analogue of eosin (which stains blood cells). In general, good perfusion and preparation of the tissue should help prevent blood vessel staining but it may not be possible to eliminate it entirely. In our experience it is generally possible to distinguish neuronal from blood vessels staining by eye.
Excitation Emission:
FJB visualization is accomplished using blue light or a 488 nm Laser. Excitation Peak: 495 nm Emission Peak: 521 nm Filter system for visualizing: Fluorescein/FITC
This product is equivalent to discontinued product AG310 (FJB) from Merck-Millipore TR-100-FTJ | TR-100-FJ | TR-160-FJC (equivalent to discontinued product AG325 (FJB) from Merck-Millipore)
Purity:
Thin layer chromatograpy using cellulose plates and a solvent system of n-propinol, water, and ammonium hydroxide (6:5:2) revealed the presence of two fluorescent isomers and two trace non-fluorescent bands. No amount of fluorescein or Fluoro-Jade was present.
The causes and effects of neuronal degeneration are of major interest to a wide variety of neuroscientists. Paralleling this growing interest is an increasing number of methods applicable to the detection of neuronal degeneration. Fluoro-Jade C stains all degenerating neurons regardless of specific insult or mechanism of cell death. Fluoro-Jade C exhibits the greatest signal to background ratio, as well as the highest resolution. This translates to a stain of maximal contrast and affinity for degenerating neurons. This makes it ideal for localising not only degenerating nerve cell bodies but also distal dendrites, axons and terminals. The dye is highly resistant to fading and is compatible with virtually all histological processing and staining protocols.
Product Type:
Staining Reagent
Format:
The reagents in the Fluoro Jade kit (10X) are all supplied in a liquid format and are ready-to-dilute.
Species Reactivity:
Human,Mouse,Other Mammals (Predicted),Rat
Applications:
FC,ICC,IHC-Frozen,IHC-Paraffin-embedded
Application Details:
The Fluoro-Jade C 'Ready to Dilute' (RTD) Staining Kit provides an easy to use assortment of Fluoro-Jade C, DAPI, sodium hydroxide and potassium permanganate in liquid form. Following our detailed protocol, Fluoro-Jade C labelled degenerating neurons are visualised with blue light excitation while DAPI counter stained cell nuclei are visualised with ultra-violet illumination. The Fluoro-Jade C Staining Kit can be used on all kinds of preserved tissues, including fresh-frozen, paraformaldehyde or formalin fixed, and formalin fixed, paraffin-embedded tissues.
Alternative Names:
FJC
Kit Components:
Materials provided: Sodium Hydroxide, Solution A (Dilute 1:10 prior to use) - 20 mL Potassium Permanganate, Solution B (Dilute 1:10 prior to use) - 20 mL Fluoro-Jade C, Solution C (Dilute 1:10 prior to use) - 20 mL DAPI, Solution D (Add to diluted Fluoro-Jade C) - 20 mL Equipment and Reagents required: Gelatin coated microscope slides Staining dishes/Coplin jars Cover slips DPX mounting media Slide warmer Convection oven Distilled water Ethanol Xylene Number of slides processed: The actual number of slides processed by this kit will depend largely upon the vessel that is used to incubate the slides. If using a standard Coplin Jar, its capacity is 50 mL and typically holds 5 slides per jar. If using such a device, then 80-100 slides stained per 50 ml of working solution (or, 5 ml of stock solution) could be processed in one day. Note the diluted dye is NOT stable and will not store overnight. It is best to use freshly diluted dye each time an experimental batch is started. Final working concentrations of FJC: 0.0001% Final working concentration of KMnO4: 0.06%
Reconstitution:
Dilute solutions as directed in the protocol instructions. Sometimes small precipitates may be present in the stock or diluted solutions. Complete mixing of the diluted solutions usually dissolves the precipitates. The precipitates, if not removed, do not usually cause any difficulties if the washing steps are followed as instructed. Optional: For entirely clean solutions Biosensis recommends filtering the diluted solution though ethanol and NaOH compatible syringe or vacuum filter devices prior to contact with tissue slides.
Storage:
The unopened kit can be stored for up to 6 months at 2-8ºC after the date of receipt. The kit and components should be stored protected from light. Diluted FJC dye solutions are not stable and should be used within 4 hours of making. The other diluted solutions can be reused and stored for up to 48 hours if refrigerated and protected from light. Best results require freshly diluted solutions. We recommend using aseptic techniques when handling the reagents to avoid bacterial growth and contamination. The FJC Ready to dilute kit is shipped ambient and stable at room temperature during transport. Refrigerate upon arrival, do not freeze.
Shelf Life:
Unopened kit 6 months at 2-8°C protected from light. See Storage instructions for working solutions recommendations.
Specificity:
Degenerating neurons, and neuronal degeneration. There is no specific staining in normal healthy brain. Note: Some researchers under some conditions report blood vessel staining with Fluoro Jade. This may be because Fluoro Jade is an analogue of eosin (which stains blood cells). In general, good perfusion and preparation of the tissue should help prevent blood vessel staining but it may not be possible to eliminate it entirely. In our experience it is generally possible to distinguish neuronal from blood vessels staining by eye.
Excitation Emission:
FJC visualization is accomplished using blue light or a 488 nm Laser. Excitation Peak: 495 nm Emission Peak: 521 nm Filter system for visualizing: Fluorescein/FITC
Fang Y et al. (2021) "Pituitary adenylate cyclase-activating polypeptide attenuates mitochondria-mediated oxidative stress and neuronal apoptosis after subarachnoid hemorrhage in rats. Free Radic Biol Med. 174:236-248. Application: IF. Species: Rat.Huan PS et al. (2021) "3,6'-Dithiopomalidomide Ameliorates Hippocampal Neurodegeneration, Microgliosis and Astrogliosis and Improves Cognitive Behaviors in Rats with a Moderate Traumatic Brain Injury." Int J Mol Sci. 22(15):8276. Application: IF. Species: Mouse.Han M et al. (2021) "Localized Modification of Water Molecule Transport After Focused Ultrasound-Induced BloodBrain Barrier Disruption in Rat Brain." Front Neurosci. 15:685977. Application: IF. Species: Rat.Gong Y et al. (2021) "Inhibition of the p?SPAK/p?NKCC1 signaling pathway protects the blood?brain barrier and reduces neuronal apoptosis in a rat model of surgical brain injury." Mol Med Rep. 24(4):717. Application: IF. Species: Rat.Zhu L et al. (2021) "Neuroprotective effects of salidroside on ageing hippocampal neurons and naturally ageing mice via the PI3K/Akt/TERT pathway." Phytother Res. [Epub ahead of print]. Application: IF. Species: Mouse.Huang Y et al. (2021) "Kisspeptin-54 attenuates oxidative stress and neuronal apoptosis in early brain injury after subarachnoid hemorrhage in rats via GPR54/ARRB2/AKT/GSK3? signaling pathway." Free Radic Biol Med. 171:99-111. Application: IF. Species: Rat.Gong Y et al. (2021) "Inhibition of the NKCC1/NF-?B Signaling Pathway Decreases Inflammation and Improves Brain Edema and Nerve Cell Apoptosis in an SBI Rat Model." Front Mol Neurosci. 14:641993. Application: IF. Species: Rat.Caron NS et al. (2021) "Mutant Huntingtin Is Cleared from the Brain via Active Mechanisms in Huntington Disease." J Neurosci. 41(4):780-796. Application: IF. Species: Human.Liu L et al. (2021) "A Novel Netrin-1-Derived Peptide Enhances Protection against Neuronal Death and Mitigates of Intracerebral Hemorrhage in Mice." Int J Mol Sci. 22(9):4829. Application: IF. Species: Mouse.Zalewska K et al. (2021) "Corticosterone Administration Alters White Matter Tract Structure and Reduces Gliosis in the Sub-Acute Phase of Experimental Stroke." Int J Mol Sci. 22(13):6693. Application: IF. Species: Mouse.Agrawal RRet al. (2021) "Neurometabolic Alterations After Traumatic Brain Injury: Links to Mitochondria-Associated ER Membranes and Alzheimers Disease." PhD Thesis. Application: IF. Species: Mouse.Zhou H et al. (2021) "AXL kinase-mediated astrocytic phagocytosis modulates outcomes of traumatic brain injury." J Neuroinflammation. 18(1):154. Application: IF. Species: Mouse.Ousta A et al. (2021) "Microglial Activation and Neurological Outcomes in a Murine Model of Cardiac Arrest." Neurocrit Care. [Epub ahead of print]. Application: IF. Species: Mouse.Horinokita H et al. (2021) "Possible involvement of progranulin in the protective effect of elastase inhibitor on cerebral ischemic injuries of neuronal and glial cells." Mol Cell Neurosci. 113:103625. Application: IF. Species: Human.Ho MH et al. (2021) "CCL5 via GPX1 activation protects hippocampal memory function after mild traumatic brain injury." Redox Biol. 46:102067. Application: IF. Species: Mouse.Lian C et al. (2021) "Pentraxin 3 secreted by human adipose-derived stem cells promotes dopaminergic neuron repair in Parkinson's disease via the inhibition of apoptosis." FASEB J. 35(7):e21748. Application: IF. Species: Mouse.Li Z et al. (2021) "The combination of deferoxamine and minocycline strengthens neuroprotective effect on acute intracerebral hemorrhage in rats." Neurol Res. [Epub ahead of print]. Application: IF. Species: Rat.Zhou K et al. (2021) "Dihydrolipoic acid enhances autophagy and alleviates neurological deficits after subarachnoid hemorrhage in rats." Exp Neurol. 342:113752. Application: IF.Asuni GP et al. (2021) "Neuronal apoptosis induced by morphine withdrawal is mediated by the p75 neurotrophin receptor." J Neurochem. [Epub ahead of print]. Application: IF.Deng S et al. (2021) "Albumin Reduces Oxidative Stress and Neuronal Apoptosis via the ERK/Nrf2/HO-1 Pathway after Intracerebral Hemorrhage in Rats." Oxid. Med. Cell. Longev. 2021, Article ID 8891373. Application: IF.Wu MY et al. (2021) "Possible mechanisms of the PERK pathway on neuronal apoptosis in a rat model of surgical brain injury." Am J Transl Res. 13(2):732-742. Application: IF.Cai G et al. (2021) "Mesenchymal stem cell-derived exosome miR-542-3p suppresses inflammation and prevents cerebral infarction." Stem Cell Res Ther. 12(1)2. Application: IF. Xu W et al. (2021) "Melanocortin 1 receptor attenuates early brain injury following subarachnoid hemorrhage by controlling mitochondrial metabolism via AMPK/SIRT1/PGC-1_ pathway in rats." Theranostics. 11(2):522-39. Application: IF. Wu M et al. (2020) "The Blood Component Iron Causes Neuronal Apoptosis Following Intracerebral Hemorrhage via the PERK Pathway." Front. Neurol. 11:588548. Application: IF. Wu D et al. (2020) "Activated WNK3 induced by intracerebral hemorrhage deteriorates brain injury maybe via WNK3/SPAK/NKCC1 pathway." Exp Neuro. 332:113386. Application: IF. Fang Y et al. (2020) "HIF-1_ Mediates TRAIL-Induced Neuronal Apoptosis via Regulating DcR1 Expression Following Traumatic Brain Injury." Front Cell Neurosci. 14:192. Application: IF. Lin CT et al. (2020) "3,6 -dithiopomalidomide reduces neural loss, inflammation, behavioral deficits in brain injury and microglial activation." Elife. 9:e54726. Application: IF. Wu H et al. (2020) "Upregulated Nmnat2 causes neuronal death and increases seizure susceptibility in temporal lobe epilepsy." Brain Res Bull. [Epub ahead of print]. Application: IF. Zahedi K et al. (2020) "Ablation of polyamine catabolic enzymes provokes Purkinje cell damage, neuroinflammation, and severe ataxia." J Neuroinflammation. 17(1):301. Application: IF. Hu X et al. (2020) "Rh-CSF1 Attenuates Oxidative Stress and Neuronal Apoptosis via the CSF1R/PLCG2/PKA/UCP2 Signaling Pathway in a Rat Model of Neonatal HIE." Oxid Med Cell Longev. 2020:6801587. Application: IF. Hu H et al. (2020) "Transient receptor potential melastatin 2 contributes to neuroinflammation and negatively regulates cognitive outcomes in a pilocarpine-induced mouse model of epilepsy." Int Immunopharmacol. 87:106824. Application: IF. Wang S et al. (2020) "Aging exacerbates impairments of cerebral blood flow autoregulation and cognition in diabetic rats." Geroscience. [Epub ahead of print]. Application: IF. Chen PY et al. (2020) "Stearic Acid Methyl Ester Affords Neuroprotection and Improves Functional Outcomes after Cardiac Arrest." Prostag Leukotr Ess. [In Press]. Application: IF. Gamdzyk M et al. (2020) "cGAS/STING Pathway Activation Contributes to Delayed Neurodegeneration in Neonatal Hypoxia-Ischemia Rat Model: Possible Involvement of LINE-1." Mol Neurobiol. 57(6): 2600-19. Application: IF. Species: Rat Tang H et al. (2020) "Delayed Recanalization After MCAO Ameliorates Ischemic Stroke by Inhibiting Apoptosis via HGF/c-met/STAT3/Bcl-2 Pathway in Rats." Exp Neurol. PMID:27527984 [Epub ahead of print] Application: IF. Species: RatDixon BJ, Chen D, Zhang Y, Flores J, Malaguit J, Nowrangi D, Zhang JH, Tang J. (2016) "Intranasal Administration of Interferon Beta Attenuates Neuronal Apoptosis via the JAK1/STAT3/BCL-2 Pathway in a Rat Model of Neonatal Hypoxic-Ischemic Encephalopathy." ASN Neuro. PMID:27683877 8(5) Application: IF. Species: RatWang JY, Huang YN, Chiu CC, Tweedie D, Luo W, Pick CG, Chou SY, Luo Y, Hoffer BJ, Greig NH, Wang JY (2016) "Pomalidomide mitigates neuronal loss, neuroinflammation, and behavioral impairments induced by traumatic brain injury in rat." J Neuroinflammation. PMID:27353053 13(1):168 Application: IF. Species: RatWang G, Manaenko A, Shao A, Ou Y, Yang P, Budbazar E, Nowrangi D, Zhang JH, Tang J (2016) "Low-density lipoprotein receptor-related protein-1 facilitates heme scavenging after intracerebral hemorrhage in mice." J Cereb Blood Flow Metab. PMID:27317656 [Epub ahead of pring] Application: IF. Species: MouseSafdaria BK, Siab TC, Wattchow DA, Smid SD (2016) "Effects of pro-inflammatory cytokines, lipopolysaccharide and COX-2 mediators on human colonic neuromuscular function and epithelial permeability." Cytokine. PMID:27177092 83:231-8 Application: IF. Species: HumanLian YD, Chen ZX, Zhu KR, Sun SY, Zhu LP (2016) "Effect of equipotent doses of bupivacaine and ropivacaine in high-fat diet fed neonatal rodent model." Brazilian Journal of Anesthesiology [In Press] Application: IF. Species: RatLaunay PS, Reboussin E, Liang H, Kessal K, Godefroy D, Rostene W, Sahel JA, Baudouin C, Parsadaniantz SM, Le Goazigo AR (2016) "Ocular inflammation induces trigeminal pain, peripheral and central neuroinflammatory mechanisms." Neurobiology of Disease. PMID:26747211 88:16-28 Application: IH. Species: MouseKhan D, Dupper A, Deshpande T, Graan PN, Steinhäuser C, Bedner P. (2016) "Experimental febrile seizures impair interastrocytic gap junction coupling in juvenile mice." J Neurosci Res. PMID:26931373 [Epub ahead of print] Application: IF. Species: MouseBye N, Christie KJ, Turbic A, Basrai HS, Turnley AM (2016) "Rho kinase inhibition following traumatic brain injury in mice promotes functional improvement and acute neuron survival but has little effect on neurogenesis, glial responses or neuroinflammation." Exp Neurol. PMID:26896832 [Epub ahead of print] Application: IH. Species: MouseBedner P, Dupper A, Hüttmann K, Müller J, Herde MK, Dublin P, Deshpande T, Schramm J, Häussler U, Haas CA, Henneberger C, Theis M, Steinhäuser C (2015) "Astrocyte uncoupling as a cause of human temporal lobe epilepsy." Brain. PMID:25765328 [Epub ahead of print] Application: IH 4%PFA, vibratome 40um sections. Species: MouseRamesh G, Didier PJ, England JD, Santana-Gould L, Doyle-Meyers LA, Martin DS, Jacobs MB, Philipp MT (2015) "Inflammation in the Pathogenesis of Lyme Neuroborreliosis." Am J Pathol. PMID:25892509 [Epub ahead of print] Application: IH with Z-fixative, 5 micron sections. Species: Rhesus MonkeySuda S, Ueda M, Nito C, Nishiyama Y, Okubo S, Abe A, Aoki J, Suzuki K, Sakamoto Y, Kimura K (2015) "Valproic acid ameliorates ischemic brain injury in hyperglycemic rats with permanent middle cerebral occlusion." Brain Res. PMID:25721785 1606:1-8 Application: IH. Species: RatMayeux JP, Teng SX, Katz PS, Gilpin NW, Molina PE (2015) "Traumatic brain injury induces neuroinflammation and neuronal degeneration that is associated with escalated alcohol self-administration in rats." Behav Brain Res. PMID:25446758 279:22-30 Application: IH. Species: RatYang LY, Chu YH, Tweedie D, Yu QS, Pick CG, Hoffer BJ, Greig NH, Wang JY (2015) "Post-trauma administration of the pifithrin-_ oxygen analog improves histological and functional outcomes after experimental traumatic brain injury." Exp Neurol. PMID:25819102 269:56-66 Application: IH. Species: RatRen YY, Zhang HQ, Duan SH, Wang SD (2015) "Influence of Dexmedetomidine on Toxicity of IntrathecalKetamine on Neonatal Rat Spinal Function." Tropical Journal of Pharmaceutical Research. 14(3) 469-77 Application: IH. Species: RatIseli CE, Merwin WH 3rd, Klatt-Cromwell C, Hutson KA, Ewend MG, Adunka OF, Fitzpatrick DC, Buchman CA (2014) "Effect of Cochlear Nerve Electrocautery on the Adult Cochlear Nucleus." Otol Neurotol. PMID: 25280052 [Epub ahead of print] Application: IH(P)Saito T, Nito C, Ueda M, Inaba T, Kamiya F, Muraga K, Katsura K, Katayama Y (2014) "Continuous oral administration of atorvastatin ameliorates brain damage after transient focal ischemia in rats." Life Sci. PMID: 24333133 94(2):106-14 Application: IH. Species: RatHayashida K, Sano M, Kamimura N, Yokota T, Suzuki M, Ohta S, Fukuda K, Hori S (2014) "Hydrogen inhalation during normoxic resuscitation improves neurological outcome in a rat model of cardiac arrest independently of targeted temperature management." Circulation. PMID: 25366995 130(24):2173-80 Application: IH. Species: Rat
The causes and effects of neuronal degeneration are of major interest to a wide variety of neuroscientists. Paralleling this growing interest is an increasing number of methods applicable to the detection of neuronal degeneration. Fluoro-Jade C stains all degenerating neurons regardless of specific insult or mechanism of cell death. Fluoro-Jade C exhibits the greatest signal to background ratio, as well as the highest resolution. This translates to a stain of maximal contrast and affinity for degenerating neurons. This makes it ideal for localising not only degenerating nerve cell bodies but also distal dendrites, axons and terminals. The dye is highly resistant to fading and is compatible with virtually all histological processing and staining protocols.
Product Type:
Staining Reagent
Format:
The reagents in the Fluoro Jade kit (10X) are all supplied in a liquid format and are ready-to-dilute.
Species Reactivity:
Human,Mouse,Other Mammals (Predicted),Rat
Applications:
FC,ICC,IHC-Frozen,IHC-Paraffin-embedded
Application Details:
The Fluoro-Jade C 'Ready to Dilute' (RTD) Staining Kit provides an easy to use assortment of Fluoro-Jade C, DAPI, sodium hydroxide and potassium permanganate in liquid form. Following our detailed protocol, Fluoro-Jade C labelled degenerating neurons are visualised with blue light excitation while DAPI counter stained cell nuclei are visualised with ultra-violet illumination. The Fluoro-Jade C Staining Kit can be used on all kinds of preserved tissues, including fresh-frozen, paraformaldehyde or formalin fixed, and formalin fixed, paraffin-embedded tissues.
Alternative Names:
FJC
Kit Components:
Materials provided: Sodium Hydroxide, Solution A (Dilute 1:10 prior to use) - 40 mL Potassium Permanganate, Solution B (Dilute 1:10 prior to use) - 40 mL Fluoro-Jade C, Solution C (Dilute 1:10 prior to use) - 40 mL DAPI, Solution D (Add to diluted Fluoro-Jade C) - 40 mL Equipment and Reagents required: Gelatin coated microscope slides Staining dishes/Coplin jars Cover slips DPX mounting media Slide warmer Convection oven Distilled water Ethanol Xylene Number of slides processed: The actual number of slides processed by this kit will depend largely upon the vessel that is used to incubate the slides. If using a standard Coplin Jar, its capacity is 50 mL and typically holds 5 slides per jar. If using such a device, then 80-100 slides stained per 50 ml of working solution (or, 5 ml of stock solution) could be processed in one day. Note the diluted dye is NOT stable and will not store overnight. It is best to use freshly diluted dye each time an experimental batch is started. Final working concentrations of FJC: 0.0001% Final working concentration of KMnO4: 0.06%
Reconstitution:
Dilute solutions as directed in the protocol instructions. Sometimes small precipitates may be present in the stock or diluted solutions. Complete mixing of the diluted solutions usually dissolves the precipitates. The precipitates, if not removed, do not usually cause any difficulties if the washing steps are followed as instructed. Optional: For entirely clean solutions Biosensis recommends filtering the diluted solution though ethanol and NaOH compatible syringe or vacuum filter devices prior to contact with tissue slides.
Storage:
The unopened kit can be stored for up to 6 months at 2-8ºC after the date of receipt. The kit and components should be stored protected from light. Diluted FJC dye solutions are not stable and should be used within 4 hours of making. The other diluted solutions can be reused and stored for up to 48 hours if refrigerated and protected from light. Best results require freshly diluted solutions. We recommend using aseptic techniques when handling the reagents to avoid bacterial growth and contamination. The FJC Ready to dilute kit is shipped ambient and stable at room temperature during transport. Refrigerate upon arrival, do not freeze.
Shelf Life:
Unopened kit 6 months at 2-8°C protected from light. See Storage instructions for working solutions recommendations.
Specificity:
Degenerating neurons, and neuronal degeneration. There is no specific staining in normal healthy brain. Note: Some researchers under some conditions report blood vessel staining with Fluoro Jade. This may be because Fluoro Jade is an analogue of eosin (which stains blood cells). In general, good perfusion and preparation of the tissue should help prevent blood vessel staining but it may not be possible to eliminate it entirely. In our experience it is generally possible to distinguish neuronal from blood vessels staining by eye.
Excitation Emission:
FJC visualization is accomplished using blue light or a 488 nm Laser. Excitation Peak: 495 nm Emission Peak: 521 nm Filter system for visualizing: Fluorescein/FITC
Fang Y et al. (2021) "Pituitary adenylate cyclase-activating polypeptide attenuates mitochondria-mediated oxidative stress and neuronal apoptosis after subarachnoid hemorrhage in rats. Free Radic Biol Med. 174:236-248. Application: IF. Species: Rat.Huan PS et al. (2021) "3,6'-Dithiopomalidomide Ameliorates Hippocampal Neurodegeneration, Microgliosis and Astrogliosis and Improves Cognitive Behaviors in Rats with a Moderate Traumatic Brain Injury." Int J Mol Sci. 22(15):8276. Application: IF. Species: Mouse.Han M et al. (2021) "Localized Modification of Water Molecule Transport After Focused Ultrasound-Induced BloodBrain Barrier Disruption in Rat Brain." Front Neurosci. 15:685977. Application: IF. Species: Rat.Gong Y et al. (2021) "Inhibition of the p?SPAK/p?NKCC1 signaling pathway protects the blood?brain barrier and reduces neuronal apoptosis in a rat model of surgical brain injury." Mol Med Rep. 24(4):717. Application: IF. Species: Rat.Zhu L et al. (2021) "Neuroprotective effects of salidroside on ageing hippocampal neurons and naturally ageing mice via the PI3K/Akt/TERT pathway." Phytother Res. [Epub ahead of print]. Application: IF. Species: Mouse.Huang Y et al. (2021) "Kisspeptin-54 attenuates oxidative stress and neuronal apoptosis in early brain injury after subarachnoid hemorrhage in rats via GPR54/ARRB2/AKT/GSK3? signaling pathway." Free Radic Biol Med. 171:99-111. Application: IF. Species: Rat.Gong Y et al. (2021) "Inhibition of the NKCC1/NF-?B Signaling Pathway Decreases Inflammation and Improves Brain Edema and Nerve Cell Apoptosis in an SBI Rat Model." Front Mol Neurosci. 14:641993. Application: IF. Species: Rat.Caron NS et al. (2021) "Mutant Huntingtin Is Cleared from the Brain via Active Mechanisms in Huntington Disease." J Neurosci. 41(4):780-796. Application: IF. Species: Human.Liu L et al. (2021) "A Novel Netrin-1-Derived Peptide Enhances Protection against Neuronal Death and Mitigates of Intracerebral Hemorrhage in Mice." Int J Mol Sci. 22(9):4829. Application: IF. Species: Mouse.Zalewska K et al. (2021) "Corticosterone Administration Alters White Matter Tract Structure and Reduces Gliosis in the Sub-Acute Phase of Experimental Stroke." Int J Mol Sci. 22(13):6693. Application: IF. Species: Mouse.Agrawal RRet al. (2021) "Neurometabolic Alterations After Traumatic Brain Injury: Links to Mitochondria-Associated ER Membranes and Alzheimers Disease." PhD Thesis. Application: IF. Species: Mouse.Zhou H et al. (2021) "AXL kinase-mediated astrocytic phagocytosis modulates outcomes of traumatic brain injury." J Neuroinflammation. 18(1):154. Application: IF. Species: Mouse.Ousta A et al. (2021) "Microglial Activation and Neurological Outcomes in a Murine Model of Cardiac Arrest." Neurocrit Care. [Epub ahead of print]. Application: IF. Species: Mouse.Horinokita H et al. (2021) "Possible involvement of progranulin in the protective effect of elastase inhibitor on cerebral ischemic injuries of neuronal and glial cells." Mol Cell Neurosci. 113:103625. Application: IF. Species: Human.Ho MH et al. (2021) "CCL5 via GPX1 activation protects hippocampal memory function after mild traumatic brain injury." Redox Biol. 46:102067. Application: IF. Species: Mouse.Lian C et al. (2021) "Pentraxin 3 secreted by human adipose-derived stem cells promotes dopaminergic neuron repair in Parkinson's disease via the inhibition of apoptosis." FASEB J. 35(7):e21748. Application: IF. Species: Mouse.Li Z et al. (2021) "The combination of deferoxamine and minocycline strengthens neuroprotective effect on acute intracerebral hemorrhage in rats." Neurol Res. [Epub ahead of print]. Application: IF. Species: Rat.Zhou K et al. (2021) "Dihydrolipoic acid enhances autophagy and alleviates neurological deficits after subarachnoid hemorrhage in rats." Exp Neurol. 342:113752. Application: IF.Asuni GP et al. (2021) "Neuronal apoptosis induced by morphine withdrawal is mediated by the p75 neurotrophin receptor." J Neurochem. [Epub ahead of print]. Application: IF.Deng S et al. (2021) "Albumin Reduces Oxidative Stress and Neuronal Apoptosis via the ERK/Nrf2/HO-1 Pathway after Intracerebral Hemorrhage in Rats." Oxid. Med. Cell. Longev. 2021, Article ID 8891373. Application: IF.Wu MY et al. (2021) "Possible mechanisms of the PERK pathway on neuronal apoptosis in a rat model of surgical brain injury." Am J Transl Res. 13(2):732-742. Application: IF.Cai G et al. (2021) "Mesenchymal stem cell-derived exosome miR-542-3p suppresses inflammation and prevents cerebral infarction." Stem Cell Res Ther. 12(1)2. Application: IF. Xu W et al. (2021) "Melanocortin 1 receptor attenuates early brain injury following subarachnoid hemorrhage by controlling mitochondrial metabolism via AMPK/SIRT1/PGC-1_ pathway in rats." Theranostics. 11(2):522-39. Application: IF. Wu M et al. (2020) "The Blood Component Iron Causes Neuronal Apoptosis Following Intracerebral Hemorrhage via the PERK Pathway." Front. Neurol. 11:588548. Application: IF. Wu D et al. (2020) "Activated WNK3 induced by intracerebral hemorrhage deteriorates brain injury maybe via WNK3/SPAK/NKCC1 pathway." Exp Neuro. 332:113386. Application: IF. Fang Y et al. (2020) "HIF-1_ Mediates TRAIL-Induced Neuronal Apoptosis via Regulating DcR1 Expression Following Traumatic Brain Injury." Front Cell Neurosci. 14:192. Application: IF. Lin CT et al. (2020) "3,6 -dithiopomalidomide reduces neural loss, inflammation, behavioral deficits in brain injury and microglial activation." Elife. 9:e54726. Application: IF. Wu H et al. (2020) "Upregulated Nmnat2 causes neuronal death and increases seizure susceptibility in temporal lobe epilepsy." Brain Res Bull. [Epub ahead of print]. Application: IF. Zahedi K et al. (2020) "Ablation of polyamine catabolic enzymes provokes Purkinje cell damage, neuroinflammation, and severe ataxia." J Neuroinflammation. 17(1):301. Application: IF. Hu X et al. (2020) "Rh-CSF1 Attenuates Oxidative Stress and Neuronal Apoptosis via the CSF1R/PLCG2/PKA/UCP2 Signaling Pathway in a Rat Model of Neonatal HIE." Oxid Med Cell Longev. 2020:6801587. Application: IF. Hu H et al. (2020) "Transient receptor potential melastatin 2 contributes to neuroinflammation and negatively regulates cognitive outcomes in a pilocarpine-induced mouse model of epilepsy." Int Immunopharmacol. 87:106824. Application: IF. Wang S et al. (2020) "Aging exacerbates impairments of cerebral blood flow autoregulation and cognition in diabetic rats." Geroscience. [Epub ahead of print]. Application: IF. Chen PY et al. (2020) "Stearic Acid Methyl Ester Affords Neuroprotection and Improves Functional Outcomes after Cardiac Arrest." Prostag Leukotr Ess. [In Press]. Application: IF. Gamdzyk M et al. (2020) "cGAS/STING Pathway Activation Contributes to Delayed Neurodegeneration in Neonatal Hypoxia-Ischemia Rat Model: Possible Involvement of LINE-1." Mol Neurobiol. 57(6): 2600-19. Application: IF. Species: Rat Tang H et al. (2020) "Delayed Recanalization After MCAO Ameliorates Ischemic Stroke by Inhibiting Apoptosis via HGF/c-met/STAT3/Bcl-2 Pathway in Rats." Exp Neurol. PMID:27527984 [Epub ahead of print] Application: IF. Species: RatDixon BJ, Chen D, Zhang Y, Flores J, Malaguit J, Nowrangi D, Zhang JH, Tang J. (2016) "Intranasal Administration of Interferon Beta Attenuates Neuronal Apoptosis via the JAK1/STAT3/BCL-2 Pathway in a Rat Model of Neonatal Hypoxic-Ischemic Encephalopathy." ASN Neuro. PMID:27683877 8(5) Application: IF. Species: RatWang JY, Huang YN, Chiu CC, Tweedie D, Luo W, Pick CG, Chou SY, Luo Y, Hoffer BJ, Greig NH, Wang JY (2016) "Pomalidomide mitigates neuronal loss, neuroinflammation, and behavioral impairments induced by traumatic brain injury in rat." J Neuroinflammation. PMID:27353053 13(1):168 Application: IF. Species: RatWang G, Manaenko A, Shao A, Ou Y, Yang P, Budbazar E, Nowrangi D, Zhang JH, Tang J (2016) "Low-density lipoprotein receptor-related protein-1 facilitates heme scavenging after intracerebral hemorrhage in mice." J Cereb Blood Flow Metab. PMID:27317656 [Epub ahead of pring] Application: IF. Species: MouseSafdaria BK, Siab TC, Wattchow DA, Smid SD (2016) "Effects of pro-inflammatory cytokines, lipopolysaccharide and COX-2 mediators on human colonic neuromuscular function and epithelial permeability." Cytokine. PMID:27177092 83:231-8 Application: IF. Species: HumanLian YD, Chen ZX, Zhu KR, Sun SY, Zhu LP (2016) "Effect of equipotent doses of bupivacaine and ropivacaine in high-fat diet fed neonatal rodent model." Brazilian Journal of Anesthesiology [In Press] Application: IF. Species: RatLaunay PS, Reboussin E, Liang H, Kessal K, Godefroy D, Rostene W, Sahel JA, Baudouin C, Parsadaniantz SM, Le Goazigo AR (2016) "Ocular inflammation induces trigeminal pain, peripheral and central neuroinflammatory mechanisms." Neurobiology of Disease. PMID:26747211 88:16-28 Application: IH. Species: MouseKhan D, Dupper A, Deshpande T, Graan PN, Steinhäuser C, Bedner P. (2016) "Experimental febrile seizures impair interastrocytic gap junction coupling in juvenile mice." J Neurosci Res. PMID:26931373 [Epub ahead of print] Application: IF. Species: MouseBye N, Christie KJ, Turbic A, Basrai HS, Turnley AM (2016) "Rho kinase inhibition following traumatic brain injury in mice promotes functional improvement and acute neuron survival but has little effect on neurogenesis, glial responses or neuroinflammation." Exp Neurol. PMID:26896832 [Epub ahead of print] Application: IH. Species: MouseBedner P, Dupper A, Hüttmann K, Müller J, Herde MK, Dublin P, Deshpande T, Schramm J, Häussler U, Haas CA, Henneberger C, Theis M, Steinhäuser C (2015) "Astrocyte uncoupling as a cause of human temporal lobe epilepsy." Brain. PMID:25765328 [Epub ahead of print] Application: IH 4%PFA, vibratome 40um sections. Species: MouseRamesh G, Didier PJ, England JD, Santana-Gould L, Doyle-Meyers LA, Martin DS, Jacobs MB, Philipp MT (2015) "Inflammation in the Pathogenesis of Lyme Neuroborreliosis." Am J Pathol. PMID:25892509 [Epub ahead of print] Application: IH with Z-fixative, 5 micron sections. Species: Rhesus MonkeySuda S, Ueda M, Nito C, Nishiyama Y, Okubo S, Abe A, Aoki J, Suzuki K, Sakamoto Y, Kimura K (2015) "Valproic acid ameliorates ischemic brain injury in hyperglycemic rats with permanent middle cerebral occlusion." Brain Res. PMID:25721785 1606:1-8 Application: IH. Species: RatMayeux JP, Teng SX, Katz PS, Gilpin NW, Molina PE (2015) "Traumatic brain injury induces neuroinflammation and neuronal degeneration that is associated with escalated alcohol self-administration in rats." Behav Brain Res. PMID:25446758 279:22-30 Application: IH. Species: RatYang LY, Chu YH, Tweedie D, Yu QS, Pick CG, Hoffer BJ, Greig NH, Wang JY (2015) "Post-trauma administration of the pifithrin-_ oxygen analog improves histological and functional outcomes after experimental traumatic brain injury." Exp Neurol. PMID:25819102 269:56-66 Application: IH. Species: RatRen YY, Zhang HQ, Duan SH, Wang SD (2015) "Influence of Dexmedetomidine on Toxicity of IntrathecalKetamine on Neonatal Rat Spinal Function." Tropical Journal of Pharmaceutical Research. 14(3) 469-77 Application: IH. Species: RatIseli CE, Merwin WH 3rd, Klatt-Cromwell C, Hutson KA, Ewend MG, Adunka OF, Fitzpatrick DC, Buchman CA (2014) "Effect of Cochlear Nerve Electrocautery on the Adult Cochlear Nucleus." Otol Neurotol. PMID: 25280052 [Epub ahead of print] Application: IH(P)Saito T, Nito C, Ueda M, Inaba T, Kamiya F, Muraga K, Katsura K, Katayama Y (2014) "Continuous oral administration of atorvastatin ameliorates brain damage after transient focal ischemia in rats." Life Sci. PMID: 24333133 94(2):106-14 Application: IH. Species: RatHayashida K, Sano M, Kamimura N, Yokota T, Suzuki M, Ohta S, Fukuda K, Hori S (2014) "Hydrogen inhalation during normoxic resuscitation improves neurological outcome in a rat model of cardiac arrest independently of targeted temperature management." Circulation. PMID: 25366995 130(24):2173-80 Application: IH. Species: Rat
Black-Gold II is a novel haloaurophosphate complex which localises myelin within the central nervous system. The Black Gold II Ready-to-Dilute (RTD) Staining Kit allows you to localise myelin, both individual fibres and tracts, along with the option of co-localising cell bodies via the Toluidine Blue counter stain. Black Gold II labelled myelinated fibres appear nearly black while the Toluidine Blue O labelled cellular Nissl bodies are blue under bright field illumination. Black Gold II can demonstrate and characterise specific myelin changes associated with exposure to diverse neurotoxicants including kainic acid, domoic acid, 3-nitropropionic acid, Fluoro-Gold and isoniazid. Black Gold II can also be combined with other histochemical markers including Nissl stains, retrogradely transported fluorescent tracers and fluorescent markers of neuronal degeneration. The advantages associated with the Black-Gold II include high resolution, high contrast, short histochemical processing time, versatility and consistent reproducibility.
Product Type:
Staining Reagent
Format:
The reagents in the Black Gold kit (10X) are all supplied in a liquid format and are ready-to-dilute.
Species Reactivity:
Mammals
Applications:
IHC-Frozen,IHC-non-Paraffin-embedded
Application Details:
Black Gold II is a high resolution myelin stain with amyloid plaque counter stain. Its use is tailored to studies using formalin or paraformaldehyde fixed, non-paraffin embedded, non-solvent processed brain tissue. It can be used with both thick and thin sections. For thick sections, gelatin coated slides or slides specially designed to bind tissues sections should be use to avoid section loss. Free-floating sections can be used as well but sections are easier to handle and transfer when mounted on slides. A suggested method for thick sections is provided as a guide: Either frozen or vibratome sections are cut at a thickness of 20-50 ?m and collected in 0.1 M neutral phosphate buffer. The sections are then typically mounted on 1% gel-coated slides and then air dried on a slide warmer (at 50°C) for at least an hour until throughly dried and adhered to the slide. The sections can be stained loose, although the sections are easier to handle when mounted on slides. The mounted sections were rehydrated in distilled water for 2 minutes before being processed in the staining solutions.
Alternative Names:
BlackGold, Black and Gold
Kit Components:
Black-Gold II (Dilute 1:10 prior to use) - 10 mL Sodium Thiosulfate, fixative (Dilute 1:10 prior to use) - 10 mL Toluidine Blue O (Dilute 1:10 prior to use) - 10 mL Acetic Acid (Dilute 1:10 prior to use) - 10 mL
Storage:
The kit can be transported at room temperature. Once received, the kit canbe stored for up to 12 months at 2-8°C protected from light. Diluted solutionscan be stored up to one month at 2-8°C protected from light.
Shelf Life:
Unopened kit 6 months at 2-8°C protected from light. See Storage instructions for working solutions recommendations.
Specificity:
Black-Gold II is a novel haloaurophosphate complex which localises myelin within the central nervous system.
Del Fiacco M et al. (2018) TRPV1-Like Immunoreactivity in the Human Locus K, a Distinct Subregion of the Cuneate Nucleus. Cells. 2018 Jul 8;7(7). pii: E72. Ying YL et al. (2014) Adult neural precursor cells from the subventricular zone contribute significantly to oligodendrocyte regeneration and remyelination. J Neurosci. 2014 Oct 15;34(42):14128-46
Research Areas:
Biosensis Stains & Tracing Reagents
Purity:
Purified
Cookies:
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