I have an antibody specific just for ICC and WB but I should use the same antibody to perform an IHC staining. Is it possible to use this ab also in IHC?
If in the datasheet it's not specified that the antibody is suitable for IHC, what it means that the manufacturer company didn't test it for IHC. You always can try and see what happens, don't forget to use a positive control (e.g. a antibody for the same protein that 100% works in IHC)
Hello Federica Sella
Having knowledge about the target protein and the epitope recognized by the antibody is important. If the target molecule is denatured namely, through fixation, reduction or pH changes, the epitope may be altered, and this may affect its ability to interact with an antibody.
IHC, ICC and Western Blot samples are processed differently. For example, ICC samples, compared to IHC, undergo a shorter fixation period. In IHC the proteins are cross linked with the fixative which will also affect the structure of the protein of interest. The antibody may not detect formaldehyde fixed antigens and excessive crosslinking of antigens may reduce the signal. In Western Blot the protein is denatured and so the antibody will recognize proteins in their denatured form.
Please note that if the antibody is monoclonal and if the datasheet specifies that it can be used for both IHC and Western Blot, it means that the antibody can bind to the linear peptide on the surface of the protein which can work for both IHC as well as Western Blot. If the antibody is polyclonal, then it will work for both IHC as well as Western Blot because it will bind linear epitopes (in case of Western Blot) and surface epitopes which could either be conformational or linear (in case of IHC).
So, please check the details in the datasheet.
Best.
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