Cytokine

Keith Bahjat kbahjat at ufl.edu
Mon Sep 11 15:39:28 EST 2000


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 at ufl.edu


> From: Jens Fleischer <jfleischer at knuut.de>
> Date: Sat, 09 Sep 2000 14:55:31 +0200
> To: Cytometry Mailing List <cytometry at 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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