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
Biosensis Brand:
Biosensis® RTD
Detection Method:
Fluorescence
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.
Shelf Life:
6 months after date of receipt (10X stock solution)
Use:
For research use only.
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.
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.
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.
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
Biosensis Brand:
Biosensis® RTD
Detection Method:
Fluorescence
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. <br>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.
Shelf Life:
6 months after date of receipt (unopened vial).
Use:
For research use only.
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
Product references:
Su IJ et al. (2021) "The Beneficial Effects of Combining Anti-A? Antibody NP106 and Curcumin Analog TML-6 on the Treatment of Alzheimer's Disease in APP/PS1 Mice." Int J Mol Sci. 23(1):556; Application: IHC/IF Species: Mouse 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: Human Hascup 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: Mouse Hascup 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
Specificity:
Amyloid plaques both intraneuronal and vascular for A-G, Etbr, nuclei and cell bodies both DNA and RNA label
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.
Purification:
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
Biosensis Brand:
Biosensis® RTD
Detection Method:
Fluorescence
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.
Shelf Life:
6 months after date of receipt (unopened vial).
Use:
For research use only.
Kit Components:
5 mL of 100X Amylo-Glo RTD (A-G RTD) solution
Product references:
Silvin A et al. (2022) "Dual ontogeny of disease-associated microglia and disease inflammatory macrophages in aging and neurodegeneration" Immunity. [Epub ahead of print]; Application: IHC/IF Species: Mouse Shrader JM et al. (2022) "Distinct Brain Proteomic Signatures in Cerebral Small Vessel Disease Rat Models of Hypertension and Cerebral Amyloid Angiopathy" J Neuropathol Exp Neurol. [Epub ahead of print]; Application: IHC/IF Species: Rat Zagorski K et al. (2022) "Immunogenicity of MultiTEP-Platform-Based Recombinant Protein Vaccine, PV-1950R, Targeting Three B-Cell Antigenic Determinants of Pathological ?-Synuclein" Int J Mol Sci. [Epub ahead of print]; Application: IHC/IF Species: Mouse Shabestari SK et al. (2022) "Absence of microglia promotes diverse pathologies and early lethality in Alzheimers disease mice" Cell Rep. 39(11):110961; Application: IHC/IF Species: Mouse Davis J et al. (2022) "rTg-D: A novel transgenic rat model of cerebral amyloid angiopathy Type-2." Cerebral Circulation - Cognition and Behavior [Epub ahead of print]; Application: IHC/IF Species: Rat Salvadores N et al. (2022) "A? oligomers trigger necroptosis-mediated neurodegeneration via microglia activation in Alzheimer's disease." Acta Neuropathol Commun. 10(1):31; Application: IHC/IF Species: Human Javonillo DI et al. (2022) "Systematic Phenotyping and Characterization of the 3xTg-AD Mouse Model of Alzheimer's Disease." Front Neurosci. 15:785276; Application: IHC/IF Species: Mouse Hohsfield LA et al. (2022) "MAC2 is a long-lasting marker of peripheral cell infiltrates into the mouse CNS after bone marrow transplantation and coronavirus infection." Glia. [Epub ahead of print]; Application: IHC/IF Species: Mouse Tsay HJ et al. (2021) "EK100 and Antrodin C Improve Brain Amyloid Pathology in APP/PS1 Transgenic Mice by Promoting Microglial and Perivascular Clearance Pathways." Int J Mol Sci. 22(19):10413; Application: IHC/IF Species: Mouse Henningfield CM et al. (2021) "Microglia-specific ApoE knock-out does not alter Alzheimer's disease plaque pathogenesis or gene expression." Glia. [Epub ahead of print]; Application: IHC/IF Species: Mouse Da Mesquita S et al. (2021) "Meningeal lymphatics affect microglia responses and anti-A? immunotherapy." Nature. 593(7858):255-260; Application: IHC/IF Species: Mouse Lauterborn 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: Human Kim JH et al. (2021) "Gamma subunit of complement component 8 is a neuroinflammation inhibitor." Brain. 144(2):528-552; Application: IHC/IF Species: Mouse Claes 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: Mouse Crapser JD. (2021) "Investigating microglial regulation of the extracellular matrix in health and neurodegenerative disease." PhD Thesis ; Application: IHC/IF Species: Human Baglietto-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: Mouse Mistrik 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: Human Lemoine 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: Human Hascup 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: Mouse Holloway 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: Mouse McQuade 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: Mouse Hascup 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: Mouse Crapser JD et al. (2020) "Microglia facilitate loss of perineuronal nets in the Alzheimer's disease brain." EBioMedicine. 58:102919; Application: IHC/IF Species: Mouse Abe 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: Mouse Zhu 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: Rat Majewski 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: Mouse Davtyan 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: Mouse Yeh 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: Mouse Bharani 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: Human Hovakimyan 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: Human Hasselmann 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: Mouse Spangenberg 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: Human Eggers 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: Rat Dominguez E (2019) "Microglial Contributions to Alzheimer's Disease Pathogenesis." PhD Thesis, UC Irvine. Application: IHC/IF Species: Mouse Jovic 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: Mouse Collins 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: Mouse Shukla 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: Mouse Feng 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, mouse Davis 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: Rat Palombo 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: Mouse Abud EM (2017) "Generation of Human Microglia from Induced Pluripotent Stem Cells to Study Innate Immunity in Neurological Diseases." PhD Thesis. 2017; Application: IF Species: Mouse Abud EM et al. (2017) "iPSC-Derived Human Microglia-like Cells to Study Neurological Diseases." Neuron. 2017; 49(2):278-93 Application: IF Species: Mouse Solomon 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: Human Xu 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: Mouse Katsouri 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: Mouse Esposito 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: Rat Marsh 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: Hu Kamphuis 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: Mouse Choi 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.
Specificity:
Amyloid plaques both intraneuronal and vascular
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.
Purification:
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.
Target:
Amyloid plaque
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