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-   -   Reusing Primary Antibody in Immunohistochemistry (http://www.molecularstation.com/forum/immunohistochemistry-forum/3571-reusing-primary-antibody-immunohistochemistry.html)

orestis 07-11-2008 10:50 PM

Reusing Primary Antibody in Immunohistochemistry
 
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 04-23-2009 10:28 PM

Re: Reusing Primary Antibody in Immunohistochemistry
 
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 08-01-2009 09:36 AM

Re: Reusing Primary Antibody in Immunohistochemistry
 
Quote:

Originally Posted by orestis (Post 8389)
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 11-12-2009 01:06 AM

Re: Reusing Primary Antibody in Immunohistochemistry
 
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 03-12-2013 09:33 PM

Re: Reusing Primary Antibody in Immunohistochemistry
 
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.


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