Date: Wed Feb 20 2008 - 09:54:32 EST
Hi Michelle, A useful control for apoptosis assays is Jurkat cells triggered with anti-Fas antibody. Briefly, anti CD95 [clone CH11] at 100 ng/ml added to Jurkat cells for 2-4 H provides an apoptotic and nonapoptotic population. For details, I refer you to: Mc Closkey, TW, Phenix BN, and Pahwa S, Flow cytometric detection and quantification of apoptotic cells, in: Manual of Molecular and Clinical Laboratory Immunology [vol. 7], B Detrick, RG Hamilton, and JD Folds, eds, Washington, DC, ASM Press, 281-290, 2006. Another recommendation would be to look at the cells under the microscope. Cells with apoptotic nuclei are typically readily visible, particularly when labeled with nucleic acid dyes. I would also recommend the use of a positive and negative control for your assay development speciifc to the cell type or cell line you are working with. And finally, using a second assay is a good idea to confirm and verify your data. Good luck, Tom Thomas W. Mc Closkey, Ph. D. Associate Director, Cellular Immunology Research and Development ICON Central Laboratory 123 Smith Street Farmingdale, NY 11735 Tel: + (1) 631-306-9789 Fax: + (1) 631-694-2524 Email: mccloskeyt@iconlabinc.com <mailto:mccloskeyt@iconlabinc.com> Web: www.icolabs.com ________________________________ From: Michelle Mok Meng Huang [mailto:michellemok@bsf.a-star.edu.sg] Sent: Monday, February 18, 2008 9:24 PM To: cyto-inbox Subject: Looking for subG1 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 ********************************************************************** This e-mail transmission may contain confidential or legally privileged information from ICON Central Laboratories, Inc. intended only for the use of the individual(s) or entity named on the transmission sheet. If you are not the intended recipient, you are here by notified that any disclosure, copying, distribution, use or taking of action in reliance upon the contents of this e-mail is strictly prohibited. If you have received this e-mail transmission in error, please notify us immediately by telephone or e-mail so that ICON Central Laboratories, Inc. can arrange for the return of the documents to us at no cost to you, and then please delete the message. Thank You, ICON Legal Department, ICON Central Laboratories, Inc. (631) 777-8833 **********************************************************************
