Cell cycle, BrdU incorporation, and Apoptosis (TUNEL)

From: Matthew Janes <mjanes@uci.edu>
Date: Wed Jun 21 2006 - 15:04:01 EDT
I want to determine in what stage of the cell cycle particular 
primary lymphocytes are dying.	So ask the community if anyone has a 
recommended protocol for CELL CYCLE (DNA content with BrdU 
incorporation to properly distinguish all cell cycle phases) with 
TUNEL, or another well established method for assaying for apoptosis.

Or can sorting of populations in the different stages of the cell 
cycle be performed?  I am experienced with sorting, so is it a 
problem sorting different populations when aquiring data on a linear 
scale (like when using PI, 7AAD, or DAPI for cell cycle?   Would the 
community recommend just sorting different lymphocytes based off 
their cell cycle, the subsequently enumerate apoptosis by microscopy, 
or look at the degree of DNA fragmentation on a gel.

Any opinion on such a task would be greatly appreciated.

best,

Matt
-- 
Matthew R. Janes, MSPH
Ph.D. Candidate
Department of Molecular Biology & Biochemistry
Center for Immunology
University of California at Irvine
mjanes@uci.edu
Received on Thu Jun 22 12:13:42 2006

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