This kit is designed to detect mouse primary antibodies that are bound to human tissue or cell antigens. Frozen or paraffin-embedded tissues, freshly prepared lymphocytes, and fixed cultured cells can be used.
Streptavidin-Biotin Amplification has been acknowledged as the premier detection method for IHC, a methodology now widely used in basic research and routine testing, providing enhanced sensitivity and performance in immunohistochemistry (IHC) applications. DAB stained slides can be dehydrated and permanently mounted using mounting medium.The kit is designed to reveal the presence of antigens in human tissue or cell preparations and can be used with all common sample preparation methods, including: frozen tissues, paraffin-embedded sections, and cell smears.
After formalin-fixed, paraffin-embedded tissue sections have been deparaffinized in xylene and dehydrated in a graded series of ethanol, an endogenous peroxidase quenching step is performed. This endogenous peroxidase activity can be quenched by either using 3% hydrogen peroxide in methanol. Non-immune serum is used to eliminate non-specific background. After a user-supplied primary antibody has been incubated with a sample, the method uses an enhanced, human-absorbed, biotinylated, affinity purified secondary antibody to react with the primary antibody. Enhanced horseradish peroxidase conjugated streptavidin (HRP-SA) is then bound to the biotinylated secondary antibody. The chromogen is then added, and the peroxidase catalyzes the substrate (hydrogen peroxide) and converts the chromogen to either a brown (DAB) or red (AEC) deposit, which visualizes the location of the antigen. This kit can also be used with automated stainers.
Kit Components:
I7506-03A: Reagent A, Serum blocking solution, 10% 1x110ml
I7506-03B: Reagent B, Second Antibody (Biotin) 1x110ml
I7506-03C: Reagent C, Streptavidin (HRP) 1x110ml
Storage and Stability:
Store all components at 4C. Stable for at least 6 months.
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