I've found substituting a 4% formaldehyde fixation in place of BD's facslyse step greatly improves performance of the permeabilizing solution. I routinely use this for cytokine analysis of both human and murine leukocyte suspensions. And this way, if you're using PBMCs, you don't have to spend all that money on the lysing solution. I recommend Polyscience's EM grade methanol free 10% formaldehyde (catalog# 04018). I get similar results as when using freshly reconstituted formaldehyde (from paraformaldhyde polymer), but this way you don't have to continue making fresh solutions all the time. In my hands, it has been stable for years in its stock 10% form, and good for a couple of weeks after diluting to 1%. kb -- Keith Bahjat Graduate Assistant University of Florida College of Medicine Gainesville, Florida Phone: (352) 392-4887 Fax: (352) 392-5393 kbahjat@ufl.edu > From: Jens Fleischer <jfleischer@knuut.de> > Date: Sat, 09 Sep 2000 14:55:31 +0200 > To: Cytometry Mailing List <cytometry@flowcyt.cyto.purdue.edu> > Subject: Re: Cytokine > > > > The BDIS anti-cytokine antibodies work with the BD Lysing Solution and the > BD Perm Solution. Even if you use PBMC and not whole blood you have to use > the Lysing solution to fix the cells before staining. In contrast, some of > the BD Pharmingen antibodies require their Pharmingen Fix and Perm > Solutions. The recognized epitopes change depending on the method of > fixation and permeabilization. I don't know which "homebrew" recipe may be > compatible with all those clones. > > Jens > > >
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