AW: BRDU DNAse

From: Wolfgang Beisker (beisker@GSF.DE)
Date: Wed Jun 16 1999 - 04:35:09 EST


Adrian,

with the DAPI staining seems strange to me. That GFP staining is whiped out
by HCL treatment, is o.k., because HCL extracts proteins.

Perhaps you can use another detection method, how long did you label the
cells and what was the concentration of BrdUrd during incubation?

Depending on that, you may be able to detect proliferation by using a
combination of PI and TO-PRO-3, excited with a HE-Ne laser and a 488nm
laser.


Best regards from Munich


Wolfgang


GSF- Flow cytometry group
Ingolstaedter Landstr. 1
85764 Munich, Germany



> -----Ursprüngliche Nachricht-----
> Von: Senderowicz, Adrian (NCI) [mailto:sendero@helix.nih.gov]
> Gesendet am: Montag, 14. Juni 1999 18:02
> An: Cytometry Mailing List
> Betreff: 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.
> DTP Clinical Trials Unit
> National Cancer Institute
> 10 Center Dr. MSC 1594
> Bldg 10, rm 6N113
> Bethesda, MD 20892
> 301-496-4119
> 301-480-7456(fax)
> sendero@helix.nih.gov
>
>
>



This archive was generated by hypermail 2b29 : Wed Apr 03 2002 - 11:53:37 EST