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Antibody validation Practical guide to finding and validating suitable antibodies for research

1.C Decide on the experimental techniques you wish to use

Many commercially available antibodies on the market have not been rigorously tested, and in general have been used for a single application, often testing reactivity with only a recombinant immunogen with no further specificity testing. Also, it is well known that the performance of an antibody in one application cannot predict its performance in another application. For example, there are many antibodies that recognise a protein that is fully denatured, but are not able to detect the same protein in its natural conformation. These antibodies would work well for Western blotting but not for immunoprecipitation or other applications that require detection of native protein and vice versa. Additionally, antibodies that recognises a formalin resistant epitope for immunohistochemistry may also work well in other techniques using formalin fixation, such as chromatin immunoprecipitation.