For in-vitro Diagnostic Use. The BrightVision two components detection system peroxidase Goat Anti-Mouse/Rabbit IgG HRP, is intended for use in immunohistochemistry for the detection of mouse or rabbit antibodies.
The BrightVision detection system peroxidase Goat Anti- Mouse/Rabbit HRP, is a Ready-to-Use system that has been manufactured to give an optimal staining, when using the protocol advised in this IFU. Prior to staining some routine fixed, paraffin-embedding tissue sections should be subjected to pre-treatment (HIER or digestive enzyme). The BrightVision detection system detects Mouse or Rabbit bound to an antigen in tissue sections. This polymer-complex is then visualized with a suitable substrate/chromogen (not provided).
Principle of method:
Two steps detection system goat anti-mouse/rabbit HRP
Reagents provided:
Post-blocking (ready-to-use) 55 ml and Polymer Goat Anti-Mouse/Rabbit HRP (ready-to-use) 55 ml.
Storage and handling:
2-8°C and in the dark
Procedure:
1. Deparaffinize and rehydrate tissue section (slide/tissue peparing), 2. Wash Aqua dest (Wash; 2x 5 min), 3. If applicable, HIER or digestive enzyme (pre-treatment), 4. Wash buffer (PBS or TBS buffer; 2x 5 min), 5. H2O2 (conc3%) (Tissue preparing; 10 min), 6. Wash buffer (PBS or TBS buffer; 2x 5 min), 7. Primary mouse or rabbit antibody (Antibody; 30 min), 8. Wash buffer (PBS or TBS buffer; 2x 5 min), 9. Detection system, step 1, post-blocking (Post-blocking; 15 min), 10. Wash buffer (PBS or TBS buffer; 2x 5 min), 11. Detection system, step 2, polymer Mouse/Rabbit HRP (Labeled polymer; 30 min), 12. Wash buffer (PBS or TBS buffer; 2x 5 min), 13. If applicable Substrate (DAB), 14. Wash aqua dest (Wash; 2x 2 min), 15. Counterstain, dehydrate and coverslip (Auxiliary)
Quality Control:
A positive control, negative control and reagent control are needed and processed in the same way as the unknow specimen slide to interpret staining results.
The aqua mounting medium is a widely used for coverslipping from aqueous solutions and is non-fluorescing and has an antifade component to increase the viewing time of the specimen. Use Aqua-Poly/Mount with most fluorescent dyes and stains including DAB, Alkaline Phosphatase, Fast Red, AEC (aminoethylcarbazole) and a variety of other chromogens to enhance and retain fluorescent intensity. The mounting medium (water based) can be used for frozen sections, fat stains, chromogens for immunohistochemistry and in situ hybridization as well as other applications requiring a water-soluble mounting medium.
Principle of method:
Coverslipping from aqueous solutions
Reagents provided:
50 ml Mounting medium (water based), ready-to-use.
Storage and handling:
Store at room temperature
Procedure:
1. Prepare slides as required. 2. Prior to coverslipping rinse the slides in distilled or deionized water. 3. Place the bottle upside down with the end cap attached in a container before use. This will help clear of tiny bubbles. 4. Blot excess water from the slide without drying the tissue specimen. The tissue must be moist prior to mounting. 5. Apply enough drops of aqua mounting medium on the tissue section so that the tissue is completely covered. 6. Carefully lower the coverslip at an angle while gently applying pressure to force any excess medium and air bubbles away from the tissue and out from under the coverslip. 7. Slides can be dried at room temperature or at 4°C or in an oven but the temperature hight is depending what type of staining is used. 8. Allow the aqua mounting medium to dry before examine under the microscope.
CryoCompound freezing medium is an aqueous based frozen tissue embedding medium designed to support tissue blocks in cryostat sectioning. Formulated to promote rapid freezing, enhanced sectioning and consistent results at a working temperature of -20° C. The ready-to-use Cryocompound is available in different colors: MON-APP900C (Transparant/colorless), MON-APP900B (Blue), MON-APP900Y (Yellow), MON-APP900O (Orange), MON-APP900G (green), MON-APP900R (Red) or Multicolorkit MON-APP900BYGR (Blue, Yellow, Green, Red).
Principle of method:
Freezing medium
Reagents provided:
100 ml Cryocompound, freezing medium (ready-to-use)
Storage and handling:
Store at room temperature
Procedure:
1. Place few drops of CryoCompound (depends on the size of the sample to be embedded) onto the center of the bottom of cryomold. Be careful to select the proper size embedding mold according to the size of the sample to be embedded. 2. Try to avoid the formation of air bubbles. Remove any bubbles inside the CryoCopound. This is important because the air bubbles will create problems when cutting sections. Air bubbles create freeze- thaw-freeze cycle and ice crystal will form inside of it and result in a very bad morphology due to the ice crystal artefact. 3. Let it settle for 15-30 seconds to allow the CryoCompound to completely wet the surface of the tissue. Hardening of the CryoCompound included the sample (it will happen in 0.5-1 minute) before preparing the CryoCompoundsample-block.
CryoCompound freezing medium is an aqueous based frozen tissue embedding medium designed to support tissue blocks in cryostat sectioning. Formulated to promote rapid freezing, enhanced sectioning and consistent results at a working temperature of -20° C. The ready-to-use Cryocompound is available in different colors: MON-APP900C (Transparant/colorless), MON-APP900B (Blue), MON-APP900Y (Yellow), MON-APP900O (Orange), MON-APP900G (green), MON-APP900R (Red) or Multicolorkit MON-APP900BYGR (Blue, Yellow, Green, Red).
Principle of method:
Freezing medium
Reagents provided:
100 ml Cryocompound, freezing medium (ready-to-use)
Storage and handling:
Store at room temperature
Procedure:
1. Place few drops of CryoCompound (depends on the size of the sample to be embedded) onto the center of the bottom of cryomold. Be careful to select the proper size embedding mold according to the size of the sample to be embedded. 2. Try to avoid the formation of air bubbles. Remove any bubbles inside the CryoCopound. This is important because the air bubbles will create problems when cutting sections. Air bubbles create freeze- thaw-freeze cycle and ice crystal will form inside of it and result in a very bad morphology due to the ice crystal artefact. 3. Let it settle for 15-30 seconds to allow the CryoCompound to completely wet the surface of the tissue. Hardening of the CryoCompound included the sample (it will happen in 0.5-1 minute) before preparing the CryoCompoundsample-block.
CryoCompound freezing medium is an aqueous based frozen tissue embedding medium designed to support tissue blocks in cryostat sectioning. Formulated to promote rapid freezing, enhanced sectioning and consistent results at a working temperature of -20° C. The ready-to-use Cryocompound is available in different colors: MON-APP900C (Transparant/colorless), MON-APP900B (Blue), MON-APP900Y (Yellow), MON-APP900O (Orange), MON-APP900G (green), MON-APP900R (Red) or Multicolorkit MON-APP900BYGR (Blue, Yellow, Green, Red).
Principle of method:
Freezing medium
Reagents provided:
100 ml Cryocompound, freezing medium (ready-to-use)
Storage and handling:
Store at room temperature
Procedure:
1. Place few drops of CryoCompound (depends on the size of the sample to be embedded) onto the center of the bottom of cryomold. Be careful to select the proper size embedding mold according to the size of the sample to be embedded. 2. Try to avoid the formation of air bubbles. Remove any bubbles inside the CryoCopound. This is important because the air bubbles will create problems when cutting sections. Air bubbles create freeze- thaw-freeze cycle and ice crystal will form inside of it and result in a very bad morphology due to the ice crystal artefact. 3. Let it settle for 15-30 seconds to allow the CryoCompound to completely wet the surface of the tissue. Hardening of the CryoCompound included the sample (it will happen in 0.5-1 minute) before preparing the CryoCompoundsample-block.
CryoCompound freezing medium is an aqueous based frozen tissue embedding medium designed to support tissue blocks in cryostat sectioning. Formulated to promote rapid freezing, enhanced sectioning and consistent results at a working temperature of -20° C. The ready-to-use Cryocompound is available in different colors: MON-APP900C (Transparant/colorless), MON-APP900B (Blue), MON-APP900Y (Yellow), MON-APP900O (Orange), MON-APP900G (green), MON-APP900R (Red) or Multicolorkit MON-APP900BYGR (Blue, Yellow, Green, Red).
Principle of method:
Freezing medium
Reagents provided:
100 ml Cryocompound, freezing medium (ready-to-use)
Storage and handling:
Store at room temperature
Procedure:
1. Place few drops of CryoCompound (depends on the size of the sample to be embedded) onto the center of the bottom of cryomold. Be careful to select the proper size embedding mold according to the size of the sample to be embedded. 2. Try to avoid the formation of air bubbles. Remove any bubbles inside the CryoCopound. This is important because the air bubbles will create problems when cutting sections. Air bubbles create freeze- thaw-freeze cycle and ice crystal will form inside of it and result in a very bad morphology due to the ice crystal artefact. 3. Let it settle for 15-30 seconds to allow the CryoCompound to completely wet the surface of the tissue. Hardening of the CryoCompound included the sample (it will happen in 0.5-1 minute) before preparing the CryoCompoundsample-block.
CryoCompound freezing medium is an aqueous based frozen tissue embedding medium designed to support tissue blocks in cryostat sectioning. Formulated to promote rapid freezing, enhanced sectioning and consistent results at a working temperature of -20° C. The ready-to-use Cryocompound is available in different colors: MON-APP900C (Transparant/colorless), MON-APP900B (Blue), MON-APP900Y (Yellow), MON-APP900O (Orange), MON-APP900G (green), MON-APP900R (Red) or Multicolorkit MON-APP900BYGR (Blue, Yellow, Green, Red).
Principle of method:
Freezing medium
Reagents provided:
100 ml Cryocompound, freezing medium (ready-to-use)
Storage and handling:
Store at room temperature
Procedure:
1. Place few drops of CryoCompound (depends on the size of the sample to be embedded) onto the center of the bottom of cryomold. Be careful to select the proper size embedding mold according to the size of the sample to be embedded. 2. Try to avoid the formation of air bubbles. Remove any bubbles inside the CryoCopound. This is important because the air bubbles will create problems when cutting sections. Air bubbles create freeze- thaw-freeze cycle and ice crystal will form inside of it and result in a very bad morphology due to the ice crystal artefact. 3. Let it settle for 15-30 seconds to allow the CryoCompound to completely wet the surface of the tissue. Hardening of the CryoCompound included the sample (it will happen in 0.5-1 minute) before preparing the CryoCompoundsample-block.
CryoCompound freezing medium is an aqueous based frozen tissue embedding medium designed to support tissue blocks in cryostat sectioning. Formulated to promote rapid freezing, enhanced sectioning and consistent results at a working temperature of -20° C. The ready-to-use Cryocompound is available in different colors: MON-APP900C (Transparant/colorless), MON-APP900B (Blue), MON-APP900Y (Yellow), MON-APP900O (Orange), MON-APP900G (green), MON-APP900R (Red) or Multicolorkit MON-APP900BYGR (Blue, Yellow, Green, Red).
Principle of method:
Freezing medium
Reagents provided:
4x 100 ml Cryocompound, freezing medium (ready-to-use)
Storage and handling:
Store at room temperature
Procedure:
1. Place few drops of CryoCompound (depends on the size of the sample to be embedded) onto the center of the bottom of cryomold. Be careful to select the proper size embedding mold according to the size of the sample to be embedded. 2. Try to avoid the formation of air bubbles. Remove any bubbles inside the CryoCopound. This is important because the air bubbles will create problems when cutting sections. Air bubbles create freeze- thaw-freeze cycle and ice crystal will form inside of it and result in a very bad morphology due to the ice crystal artefact. 3. Let it settle for 15-30 seconds to allow the CryoCompound to completely wet the surface of the tissue. Hardening of the CryoCompound included the sample (it will happen in 0.5-1 minute) before preparing the CryoCompoundsample-block.
CryoCompound freezing medium is an aqueous based frozen tissue embedding medium designed to support tissue blocks in cryostat sectioning. Formulated to promote rapid freezing, enhanced sectioning and consistent results at a working temperature of -20° C. The ready-to-use Cryocompound is available in different colors: MON-APP900C (Transparant/colorless), MON-APP900B (Blue), MON-APP900Y (Yellow), MON-APP900O (Orange), MON-APP900G (green), MON-APP900R (Red) or Multicolorkit MON-APP900BYGR (Blue, Yellow, Green, Red).
Principle of method:
Freezing medium
Reagents provided:
100 ml Cryocompound, freezing medium (ready-to-use)
Storage and handling:
Store at room temperature
Procedure:
1. Place few drops of CryoCompound (depends on the size of the sample to be embedded) onto the center of the bottom of cryomold. Be careful to select the proper size embedding mold according to the size of the sample to be embedded. 2. Try to avoid the formation of air bubbles. Remove any bubbles inside the CryoCopound. This is important because the air bubbles will create problems when cutting sections. Air bubbles create freeze- thaw-freeze cycle and ice crystal will form inside of it and result in a very bad morphology due to the ice crystal artefact. 3. Let it settle for 15-30 seconds to allow the CryoCompound to completely wet the surface of the tissue. Hardening of the CryoCompound included the sample (it will happen in 0.5-1 minute) before preparing the CryoCompoundsample-block.
Paraformaldehyde (PFA) is the polymerization product of formaldehyde with a typical degree of polymerization of 8100 units. Paraformaldehyde is not a fixative itself; it must be depolymerized to formaldehyde in solution. In cell culture, a typical formaldehyde fixing procedure would involve using a 4% formaldehyde solution in phosphate buffered saline (PBS) on ice for 10 minutes. Fixing ensures that sample cell structures stay intact and that antigens are immobilized, while ideally still permitting unfettered access of antibodies to target antigens. CAS 30525-89-4, soluble in water, pH 7.0-7.6 at 25°C. The solution should be clear, colorless, with no precipitate.
Principle of method:
Fixing cells for immunohistochemistry (IHC)
Reagents provided:
500 ml 4 % Paraformaldehyde (PFA) solution in PBS
Storage and handling:
Store at -20°C for one year
Trouble shooting:
Adjust pH to 7.2-7.6 with NaH2PO4 if pH
Precautions:
Causes serious eye damage. Suspected of causing cancer. May cause an allergic skin reaction. Causes skin irritation. Avoid breathing dust/fume/gas/mist/vapors/spray. Wear protective gloves/protective clothing/eye protection/face protection. Use personal protective equipment as required. IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do. Continue rinsing. Store locked up. Dispose of contents/container in accordance with local/regional/national/international regulations
Heat-shock protein 70 (Hsp70) is the major stress-inducible protein in vertebrates and is highly conserved throughout evolution. It plays a role as a molecular chaperone and is important for allowing cells to cope with acute stressor insult, especially those affecting the protein machinery. Heat shock cognate protein 70 (HSC70), is a highly conserved protein and a member of the family of molecular chaperones.Source of HSP70 standard: recombinant Hsp70 of Arabidopsis thaliana UniProt: Q9LHA8, TAIR: AT3G12580
Product Type:
Antibody
Format:
Lyophilized in glycerol
Storage Temp:
Store lyophilized/reconstituted at -20 °C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
Concentration: after re-constitution with sterile milliQ water final concentration of the standard is 0.15 pmoles/ lProtein standard buffer composition: Glycerol 10%, Tris Base 141 mM, Tris HCl 106 mM, LDS 2%, EDTA 0.51 mM, SERVA Blue G250 0.22 mM, Phenol Red 0.175 mM, pH 8.5, 0.1mg/ml PefaBloc protease inhibitor (Roche), 50 mM DTT.This standard is ready-to-load and does not require any additions or heating. It needs to be fully thawed and thoroughly mixed prior to using. Avoid vigorous vortexing, as buffers contain detergent. Following mixing, briefly pulse in a microcentrifuge to collect material from cap.This standard is stabilized and ready and does not require heating before loading on the gel. Please note that this product contains 10% glycerol and might appear as liquid but is provided lyophilized. Allow the product several minutes to solubilize after adding water. Mix thoroughly but gently Take extra care to mix thoroughly before each use, as the proteins tend to settle with the more dense layer after freezing.
Application Details:
Standard curve: 3 loads are recommended (eg 0.1, 0.2, 0.3 pmol). Adjust range to fit your samples and your experiment. For most applications a sample load of 0.2 μg of chlorophyll/well will give a HSP70 signal in this range.Positive control: a 2 μl load per well is optimal for most chemiluminescent detection systems. Higher standard concentration needs to be used to allow detection by Coomasie stains. Such gels with higher standard concentration can not be used for quantitation using chemiluminescence.This standard is stabilized and ready and does not require heating before loading on the gel.Please note that this product contains 10% glycerol and might appear as liquid but is provided lyophilized. Allow the product several minutes to solubilize after adding water. Mix thoroughly but gently. Take extra care to mix thoroughly before each use, as the proteins tend to settle with the more dense layer after freezing.
Reconstitution:
For reconstitution add 82 l of steril water. Please notice that this product contains glycerol and might appear as liquid but is provided lyophilized. Final standard concentration is going to be 0.15 pmol/ l
Molecular Weight:
70 kDa
Special application note:
The HSP70 protein standard can be used in a combination with Agrisera global antibiodies (AS08 371) to quantitate HSP70 from a wide range of species. Global antibodies are raised against highly conserved amino acid sequence. Quantitative western blot: detailed method description, video tutorial
Research area:
Photosynthesis protein standards-quantitation
Code:
Photo20
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