Take the 2-minute tour ×
Biology Stack Exchange is a question and answer site for biology researchers, academics, and students. It's 100% free, no registration required.

Commercial suppliers of primary antibodies for given protein targets typically list recommended applications, for which the antibody has presumably been shown to work. I am usually looking to use them for immunofluorescence, and my impression (which admittedly may be based on an unrepresentative sample) is that most antibodies are recommended only for use with Western Blotting.

Why is this? Is WB the most common application? The most general? Is it easier to test for than IF?

And, perhaps more to the point, how seriously should one take these recommendations? If an antibody is not recommended for IF, does that mean that it probably won't work? Or that it may well but no-one's bothered to check? Or that it almost certainly will?

share|improve this question

2 Answers 2

up vote 6 down vote accepted

Maybe WB is easier for them to test?

In general, however, my recommendation is to check if they provide refs to paper using those antibodies and look at the images on the paper.

Be careful putting your trust in the supplier's pictures, I have seen obviously photoshopped images on commercial websites. Also always double-check the referenced papers, as sometimes they don't really use the same product that is sold...

Other than that, in my experience, the recommended application is just that: recommended. The antibody may as well work for other application. That said, note that there is a rationale for an antibody working for WB and not IHC, for instance: perfusion with fixators such as PFA will change the 3D structure of the target protein and may mask (or reveal) certain epitops, so the antibody may be more or less efficient depending what form of the protein you're dealing with (native, fixed in PFA, fixed in glutaraldehyde, SDS-denaturated etc.)

share|improve this answer

Antibodies for WB and for IF have fundamentally different requirements for the epitope structure. Antibodies for WB recognize denatured structures extended by SDS treatment. Antibodies for IF often will need to recognize a native-like structure.

Depending on the supplier, some will have lots of antibodies for IF. I suggest looking around some more.

share|improve this answer

Your Answer

 
discard

By posting your answer, you agree to the privacy policy and terms of service.

Not the answer you're looking for? Browse other questions tagged or ask your own question.