BrdU staining and surface antigen

DARZYNKIEWICZ ZBIGNIEW Z_DARZYNKIEWICZ at nymc.edu
Tue Jun 8 10:12:42 EST 2004


The "Absolute-S" kit was developed based on the SBIP (DNA Strand
Break-Induction by Photolysis) methodology described e.g in  Li et al.,
Exp. Cell Res. 222;28-37,1997; Int. J. Oncol., 4: 1157-61, or Juan et
al., Cancer Res., 57: 803-807, 1997. In this procedure there is no DNA
degradation at all - the incorporated BrdU sensitizes DNA to undergo
photolysis. Following the photolysis induced by UV light, (which may be
enhanced by cell treatment with Hoechst 33342) the resulting DNA strand
breaks are labeled with the TUNEL-like approach. Cell morphology and
expression of intracellular and surface antigens are well preserved.   

 

Zbigniew Darzynkiewicz, M.D., Ph.D.

Director, Brander Cancer Research Institute at NYMC

 

-----Original Message-----
From: Sandra Aresta [mailto:saresta at hybrigenics.fr] 
Sent: Monday, June 07, 2004 7:48 AM
To: cyto-inbox
Subject: BrdU staining and surface antigen

 

Hello everybody!

I'm trying to measure cell proliferation by flow cytometry in cells
transfected with a membrane protein. However, I cannot denature my DNA
to expose BrdU with HCl, because I loose the membrane staining. I've
tried to use DNase instead but it has been difficult to optimise (the
DNA gets too degraded, even with low amounts of DNase). Could anyone
suggest a solution?

I've seen that Phoenix Flow Systems sells a kit that should enable to
combine BrdU and other antigens detection, named "Absolute-S". There is
another kit from BD, "BrdU Flow kit", that is supposed to work as well.
Does anyone know these, and if they work OK?

Thank you in advance for your help.

Sandra

 

 

 

 

 

Sandra Aresta, PhD

Biotechnology

Hybrigenics SA

3-5 impasse Reille

75014 Paris

France

Tel: +33 (0) 1 58 10 38 00 / 60

Fax: +33 (0) 1 58 10 38 49

 

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