Hi Heather, CD69 is an early activation marker. Its expression can be detected as early as 4 hours post-stimulation and is down-regulated after 2 days. The peak of CD25 (IL-2 receptor a chain) expression is 2-4 days after activation. CD71(transferrin receptor) is expressed on proliferating cells and erythroid progenitor cells. The peak expression is between 2-5 days. I find CD69 is the best activation marker for this purpose, since staining intracellularly for most cytokine should be performed during short stimulations. For detailed stimulation protocols and intracellular flow stainining protocols for cytokine expression: http://www.ebioscience.com/ebioscience/appls/FCI.htm http://www.ebioscience.com/ebioscience/appls/BAC.htm Best wishes, Lily Lily Lai eBioscience http://www.ebioscience.com ----- Original Message ----- From: Heather Medbury <Heather_Medbury@wsahs.nsw.gov.au> To: cyto-inbox Sent: Tuesday, May 08, 2001 8:22 PM Subject: which activation marker for leukocytes > > Hi FLOWers, > > We are doing mixed lymphocyte reactions and want to detect the activated T cells by > flow cytometry, so we can examine them for cytokine production. > > asking around people have suggested we stain for either CD25, CD69 or CD71. > > I am not familiar with these markers, do they appear early or late, after how many > hours etc. > and any suggestions on the best time points to look for cytokines. > > > > Heather > > > > Heather Medbury (PhD) > Department of Surgery > Westmead Hospital > Westmead 2145 > 61-2 9845 7677 > -------------------------------------------------------------- > All experiments are preliminary > > ------------------------o0)(0o-------------------------------------- > > "In his heart a man plans his course > but the Lord determines his steps" > > --------------------------------------------------------------- >
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