RE: Looking for subG1

Our favorite cells that generate distinct "sub-G1" peak are HL-60 or
Jurkat cells treated with 0.2 uM camptothecin for 3-5 h. I want to
stress, however, that the presence of sub-G1 population, by itself,
cannot be considered the evidence of apoptosis. Thgis is a nonspecific
marker and often mechanically damaged cells even necrotic cells may end
up with fractional DNA content. Furthermore some cell types undergo
apoptosis (primarily solid tumors of different lineage) without
extensive DNA fragmentation. In such instances no sub-G1 population can
be detected. We addressed limitations, pitfalls and other problems in
detecting apoptosis by cytometry in separate article: "Difficulties and
pitfalls in analysis of apoptosis". In: Methods in Cell Biology Vol. 63,
CYTOMETRY, 3rd Edition. Academic Press, San Diego, CA, 2001; pp 527-559.


 

 

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

Professor of Pathology and Medicine 
Director, Brander Cancer Research Institute 
New York Medical College 
BSB, Room 438 
Valhalla, N.Y. 10595 

www.darzynkiewicz.com/zbigniew/ 

-----Original Message-----
From: Gerstein, Rachel [mailto:Rachel.Gerstein@umassmed.edu] 
Sent: Wednesday, February 20, 2008 3:11 PM
To: cyto-inbox
Subject: RE: Looking for subG1

 

""gold-standard" cell line know for exhibiting a sub-G1 population under
a certain treatment protocol"

IL-3 or IL-7 dependent cell lines, under cytokine withdrawal
293 cells treated with TNF-alpha
fresh mouse thymocytes, treated with corticosteroid

for subG1, be sure you are not excluding them in your FSC threshold
setting

=======================================================
Rachel M. Gerstein, Ph.D.
Associate Professor
Department of Molecular Genetics and Microbiology
Graduate Program in Immunology/Virology
University of Massachusetts Medical School
55 Lake Avenue North
Worcester, MA 01655-0002
(508) 856-1044
(508) 856-5920 (FAX)



-----Original Message-----
From: Michelle Mok Meng Huang [mailto:michellemok@bsf.a-star.edu.sg]
Sent: Mon 2/18/2008 9:24 PM
To: cyto-inbox

Dear flowers,



I have a customer here performing analysis on my LSR II looking for a
sub-G1 population. Trouble is, we haven't seen any. So we're not sure if
his treatment protocol has an issue (still in optimization) or that the
cells really do not have a sub-G1 population. So is there is a
"gold-standard" cell line know for exhibiting a sub-G1 population under
a certain treatment protocol?



Any help would be most appreciated.



Thank you,



Michelle Mok

Research Officer

Flow Cytometry