![]() If the antibody has been validated* for IHC, then the manufacturer’s guidelines or any published references can be used as guidance for the experiment. Not all antibodies work for all applications. This procedure is accomplished either by using fluorophore-conjugated primary antibodies targeting a specific protein, or by first labeling with primary antibodies followed by secondary antibody detection. IHC can also employ fluorescent means of detection. Most often, the enzymes used are horseradish peroxidase (HRP) or alkaline phosphatase (AP), which are conjugated to primary or secondary antibodies. Chromogenic detection is based on antibodies conjugated to enzymes where the staining process exploits enzymes which catalyze the deposition of a colored staining product at antigenic sites within the sample. IHC target antigens are detected directly through either chromogenic or fluorescent means, and the type of readout depends on the experimental design. Detecting the target antigen with antibodies is a multi-step process that requires optimization at every level to maximize signal detection. IHC makes it possible to visualize and document the high-resolution distribution and localization of specific cellular components within cells and within their proper histological context. Immunohistochemistry (IHC) combines anatomical, immunological, and biochemical techniques to image discrete components in tissues by using appropriately labeled antibodies to bind specifically to their target antigens in situ. ![]()
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