Posted 06 October 2011 - 09:21 PM
Several antibodies require staining at 37 degrees (chemokine receptors) or eventually RT (tetramers) and I noticed doing that, that most of the time every other markers in my panels look brighter and 'cleaner'.
I've always been told that phenotypic stains have to be performed at 4 degrees one reason for that being to limit cell death. From my experience, staining at 37 degrees or RT for no longer than 30 minutes, does not affect cell viability or only marginally (looking at pretty much every cell populations in mouse spleens, LN, BM, liver). So I was wondering if there was other rational reasons underlying why stains have to performed at +4 ?
Posted 07 October 2011 - 01:19 AM
Posted 28 November 2011 - 08:02 PM
Posted 29 November 2011 - 01:12 PM
Also, if you've fixed your cells, all cells are somewhat "dead".