Immunohistochemistry Detection Kit, Streptavidin, Rabbit, BioAssay(TM) is designed to reveal antigens that have reacted with a user-supplied primary antibody on human tissue or cell samples. 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.
I7506-04A: Reagent A, 1x110ml. Ready-to-use serum blocking solution,10% goat non-immune serum
I7506-04B: Reagent B. 1x110ml. Ready-to-use biotinylated second antibody
I7506-04C: Reagent C. 1x110ml. Ready-to-use streptavidin-peroxidase (HRP)
Materials required but not provided:
Xylene, ethanol, and absolute methanol
Distilled or deionized water
30% hydrogen peroxide
10mM phosphate-buffered saline (PBS), pH 7.4 or 50mM Tris-buffer saline (TBS), pH 7.8.
0.05% Tween 20.
Mini PAP Pen
HRP Substrate/Chromogen reagents:
*AEC Single Solution or DAB Kit
*This kit does not contain a chromogen. AEC Single Solution or DAB Kit are recommended
to be used with this kit.