I am staining some brain sections stored in cryoprotectant that express a Histone H2B- GFP fusion protein that were generated ~10 years ago. I know I need to enhance signal with an anti-GFP antibody, but do I need any specialised H2B-GFP specific primary antibody? Or will anti-GFP do the trick?
Sections are free-floating 30um stored in cryoprotectant, initially fixed in PFA.
Dear Jemima Becker ,
an anti-GFP antibody will do the trick. Since the epitope recognized by antibodys is only a few aminoacids long a anti-GFP antibody will work fine.
You should also consider that a monoclonal antibody would only bind to one epitope on your antigen (i.e. GFP) 1:1 binding. When you would use a polyclonal antibody that one is purified from a bunch of different antibodys against the antigen. Meaning that you would enhance your signal since the binding stichometry is different since the mixture of anti-GFP antibodys will bind to different epitopes on your GFP.
Best wishes Soenke
Sönke Weinert thank you so much for your help, it is greatly appreciated!
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