20 mM Tris-HCl, pH 8.0, containing 20 mg/ml BSA and 0.05% NaN3.
Availability
Shipped within 5-10 working days.
Note
THIS PRODUCT IS FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC, THERAPEUTIC OR COSMETIC PROCEDURES. NOT FOR HUMAN OR ANIMAL CONSUMPTION.
Directions for use
Suggested IHC-P Protocol
Preparation of Tris-EDTA Buffer (10 mM Tris Base, 1 mM EDTA solution, 0.05% Tween-20, pH 9.0): Mix 1.21 g Tris and 0.37 g EDTA and dissolve in 700 ml of distilled water. Adjust pH to 9.0 with 1 M HCl and then add 0.5 ml of Tween-20 and mix thoroughly. Adjust the final volume to 1 L with distilled water. Store this solution at room temperature for up to 3 months or at 4 °C for long-term storage.
Preparation of Wash Buffer: Use 0.05 M Tris-HCl, pH 7.6, containing 0.2% Tween-20.
Deparaffinize the section in 3 changes of xylene, 5 minutes each.
Wash the section in 96%, 80% and 70% ethanol, 10 minutes each.
Rinse in distilled water.
Block endogenous peroxidase by incubating the tissue in 3% hydrogen peroxide (H2O2) for 10 minutes.
Wash in distilled water.
Antigen retrieval: immerse the slide in Tris-EDTA buffer, pH 9.0, and incubate in a water bath for 30 minutes at 95 °C.
Remove the slide from the water bath and allow to stand at room temperature (in Tris-EDTA buffer, pH 9.0) for 15 minutes.
Rinse in distilled water.
Wash in Wash Buffer for 5 minutes.
Incubate the section with primary antibody at 1/100 - 1/200 dilution for 1 hour in a closed wet chamber. It is recommended to use abx291502 Primary Antibody Diluent or a diluent containing protease-free BSA (> 1 mg/ml) to dilute this antibody.
Wash twice with Wash Buffer, 5 minutes each.
Add the secondary antibody and proceed to standard immunohistochemistry protocol (HRP - Peroxide - DAB). It is recommended to use abx291501 Rabbit and Mouse HRP/DAB Detection Kit.
Wash twice in Wash Buffer, 5 minutes each.
Apply the DAB chromagen for 1-3 minutes.
Wash in distilled water for 10 minutes.
Stain in hematoxylin for 5 minutes.
Wash in distilled water for 10 minutes.
Dehydrate the section in 2 changes of 96% ethanol, 5 minutes each.
Wash the section in 2 changes of xylene, 2 minutes each.
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