dkane at amgen.com
Thu Oct 19 19:38:02 EST 1995
york at mbcrr.dfci.harvard.edu (Ian A. York) wrote:
>I'd like to lyse cells with digitonin, but I'm not clear on the
>preparation of the digitonin itself. I note that some people use Aldrich
>digitonin, and go through a fairly elaborate preparation (either
>recrystallization, or preparation one week in advance followed by letting
>it sit and filtering out crystals, I think). Others don't make any
>comments about special preparation. I note that Sigma sells two forms of
>digitonin - one is noted 'for aqueous preparations'.
>Does anyone have experience with digitonin? Comments on preparation,
Sure. I use Sigma digitonin, made in DMSO to 5mM (35mg/5.7ml DMSO),
then dilute this 10-fold in PBS, and that is a 10x stock for killing
your cells. However, cells will vary in their response, so your cells
may need more or less, depending on how long you want to wait for lysis.
My purpose in using this concentration was simply to kill monolayers
of cells without them detaching (as will happen with higher concentrations) so that I could quantify my total cell population
using a fluorescent dye.
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