Check with the manufacturer...it will depend on the precise nature of the antigen used to generate the antibody. Sometimes peptide generated antibodies only recognise the unfolded conformation as found in proteins prepared for Western blot. Failing any manufacture recommendation, do a series of serial dilutions using a known positive and known negative substrate samples to determine the specificity and sensitivity of the antibody. Use non-test tissue sections (rather than your experimental tissue-you don't want to waste that!)

It depends on presence of secondary Abs and method of visualization , eg fixed cells or sections.

Refere to manufacture r

Hi Argha,

prinicpally you can use antibodies (ab) which work for Western blots also in immunohistochemistry, but usually you need a much higher concentration of the ab in IHC. In some of our work, where the dilution of ab for WB was 1 : 10000, the necessary dilution for IHC was 1 : 200.

Regards, Daniela

to address this part of your question and support what other researchers have already said....

"....if all primary Ab for western can't be used for immunocytochemistry, what is the reason behind it?"

As Caroline mentioned, whether a primary antibody can be used for both immunocytochemistry and western blotting depends on the epitope that the antibody recognizes. More specifically, it will depend on whether that epitope changes between the protein's native folded state (as would be the case for immunoctyochemistry) and its denatured state (as would be the case for western blot).

If your primary antibody works for western but not for immunocytochemistry, it could be that the peptide region that is recognized by the antibody is inaccessible when the protein is folded. This would more likely occur for antibodies that are produced using a synthetic peptide as the antigen (rather than native protein). Not a hard and fast rule, but it could happen.

Or, it could be as simple as the way you went about preparing your ICC sample (fixative, permeabilization, etc).

Daniela's response about concentration is important as well. Antibody concentrations required for IHC/ICC are generally much higher than what is required for WB.

I would recommend starting with an antibody that has already been tested and shown to work in my desired application(s), when that is an option. Even if it's a little bit more expensive up front for that particular antibody - the time & energy you spend testing something in an unknown application could quickly become even more expensive.

Hope that helps; Best of luck to you!

you can use the antibodies but be sure that you are not getting any false positive results. manufacturer, generally validated these antibodies for only one or two or more techniques.

you have the both possibilities of getting result and not getting result.

i will suggest you to use that antibodies which are already validated for your technique.

you can purchase ab from any manufacturer, sometimes very cheap antibodies works well and some times very costly ab not work.....