[Cytometry] apoptosis posiitve control

McCloskey, Thomas Thomas.McCloskey at iconplc.com
Wed Oct 27 09:30:40 EDT 2010


Hello Alison,

The following basic procedure produces an apoptotic population:

1]  Maintain Jurkat cells in RPMI 1640 medium with 10% FCS, 2mM
L-glutamine.  
2]  Set up 12 x 75 mm tubes with 10e6 cells/ml
3]  Add anti-CD95 [100 ng/ml] antibody and incubate for 2 to 4 H at
37oC.

 CD95 antibody - clone CH-11

More details are available in:
Mc Closkey et al, Flow Cytometric Detection and Quantification of
Apoptotic Cells which is in the Manual of Molecular and Clinical
Laboratory Immunology, 2006.  

Good luck,
Tom

Thomas W. Mc Closkey, Ph. D.
Associate Director, R & D
ICON Central Laboratories 
123 Smith Street
Farmingdale, New York 11735

Tel:           + (1) 631-306-9789
Fax:          + (1) 631-694-2524
Email:       Thomas.McCloskey at iconplc.com
Web:         www.iconplc.com


-----Original Message-----
From: cytometry-bounces at lists.purdue.edu
[mailto:cytometry-bounces at lists.purdue.edu] On Behalf Of Stickney,
Alison
Sent: Tuesday, October 26, 2010 5:07 PM
To: 'cytometry at lists.purdue.edu'
Subject: [Cytometry] apoptosis posiitve control

Hi all,
I am trying to establish a positive control for an apoptosis assay using
annexin-V and 7-AAD. So far i have been using Camptothecin, but have
only been getting small numbers of apoptotic cells.  Does anyone have a
good protocol for apoptosis induction using camptothecin? i.e.
concentration, time period of incubation etc.
Regards,
Alison

Alison Stickney BVSc(hons) MVS MACVSc
Small Animal Medicine Resident
Institute of Veterinary, Animal and Biomedical Sciences
Massey University
Palmerston North 4442
New Zealand
Phone: +64 6 350 5329
Fax: +64 6 350 5616
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