Re: BRDU DNAse

From: C. Kevin Becker (ckb@phnxflow.com)
Date: Thu Jun 17 1999 - 12:54:40 EST


Subject: re: BRDU DNAse
Dear flowers,
>I am transfecting cells with GFP and when I try to do BRDU
Immunefluouresnce
>with the HCl protocol, all DAPI and GFP staining dissappears... I tried
twice
>the DNase method without any luck. Can you help?
>thanks a lot
>
>Adrian Senderowicz, M.D.

I would suggest you try formaldehyde fixation of your GFP stained cells,
digatonin permeabilization, the SBIP methodology (our Absolute-S kit for
measuring cell proliferation)substituting our PE conjugated anti-BrdU
antibody and then staining total DNA content with 7-AAD.  No Acid, No Heat,
No DNAse
   ______   ______  _______
  /_____/  /_____   /______   C. Kevin Becker
/            /             ______/ Phoenix Flow Systems, Inc.
11575 Sorrento Vlly. Rd. #208, San Diego, CA 92121 USA
(619) 453-5095 FAX (619) 259-5268 ckb@phnxflow.com



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