Reusing Primary Antibody in Immunohistochemistry

[orestis]

Hello, can we also reuse primary antibodies in immunohistochemistry IHC? I know that we can reuse primaries after westerns, but i would like to reuse the IHC antibodies to save money. Does anyone have experience in this?
thanks

[radam2k]

Normally we draw from the concentrated antibody vial only what we need for the procedure at hand. We prepare the working dilution from this draw, thus leaving the concentrated antibody in the original vial for further use. We do not re-use the dliuted antibody that has incubated with the test tissue.

[Hunydd]

Originally Posted by orestis

Hello, can we also reuse primary antibodies in immunohistochemistry IHC? I know that we can reuse primaries after westerns, but i would like to reuse the IHC antibodies to save money. Does anyone have experience in this?
thanks

Hi radam. Maybe you already solved the problem. We prepare dilutions for about 20-30 samples, with antibodies we use everyday, and they work for at least 1 month if you keep it in the fridge, and avoiding high temperatures, sunlight and a long time exposure to air oxidation. We also alicuotate dilutions for some special antibodies and keep them at -20 degrees.

[airie]

yes i agree with radam and hunydd because after incubating the tissue sections with primary antibody, the concentration of antibody left in the solution will be different, i.e lower, and it may or may not give you consistent staining quality!

[haisond]

Airie is correct, with each reuse, the antibody concentration decreases due losses by binding of antibodies to target antigens. If you still get good results with a reuse, that means that your original primary ab dilution was very concentrated. With primary antibodies there are no ways to stretch the buck here with sacrificing quality.