I am using 7-10 um frozen sections of mouse brain for IF. My target antigens are both intracellular (trying to determine) and membranous. Do I need to use any permeabilizer ? If so, which one would be the best------ Saponin /Triton-X ?
I greatly appreciate any suggestions...
Hi Sharif. You will need to permeabilise in order to detect intracellular antigens. If you are not sure about location, you can try both without and with permeabilization and then locate the antigens. I have Used of triton x 100 and it works well.
Good luck
I did surface staining on same slides in 3 color confocal microscopy. It didn't affect surface staining at all. We used PBS with 0.1% Triton-X 100 for 30 minutes.
Good luck
Usually i used NET Gel for 1h for permeabilization intracellular antigen, somtimes we used PBS with 0.1% Triton-X 100 for 30 min .
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