immunocytochemistry - (Sep/05/2007 )
Hello!!
I am interested in immunohistochemistry assays. I am looking for diferent fixation protocols and I don`t kwon what is the most aproppiated to work with an adherent cell line. In one hand, some people use 2% paraformaldehyde, mean while others do that with 3-4% paraformaldehyde during 15 mins. 
which one is the best?? Thanks.
anywhere from 2% - 4% should be fine (i use 4% for ten min on cells or frozen tissue)
dom
Some cell lines are a picky. So you might find 4% to work better. I've personally have had good luck with a 2% pfa on adherent neuronal cells.
I am interested in immunohistochemistry assays. I am looking for diferent fixation protocols and I don`t kwon what is the most aproppiated to work with an adherent cell line. In one hand, some people use 2% paraformaldehyde, mean while others do that with 3-4% paraformaldehyde during 15 mins.
which one is the best?? Thanks.some antibodies dislike paraformaldehyde-fixed cells; alternatively use -20°C cold 100% MeOH or 50:50 MeOH:aceton for 4 min; the best is to check the various agents and conc
Bearer is right. Many antibodies can be finicky about the fixation used and won't work (or give high background) with certain methods. You may need to fix your cells with a few different methods and see which works best. Also, you need to take into consideration what you want to image. Different fixations are designed to maintain different cellular morphologies and, depending on which method you use, you may disrupt what you want to see. Do some internet searches and it's pretty easy to figure out what cellular components can and can not be preserved with the various fixations.