Dear Flowers, Hello and best wishes. I want to develop an assay for intracellular IL4 production in a human T cell line. Currently I am using a Jurkat cell line. I have some doubts regarding this cell line and would be grateful for any help in resolving these issues. 1. There are a couple of Jurkat cell lines available from ATCC where it is mentioned that they produce IL2. I would like to know if any one has used any of these cell lines for looking at intracellular levels of IL4 after stimulation with PMA and Ionomycin? JurkatE6.1? 2. What are the cell surface markers which can define these cells? CD4, CD8 -ve is mentioned but what about IL2R, IL4R ? 3. Are there any other human cell lines available which can be used for detection of intracellular IL4, IL5 upon stimulationwith Ca2+ ionophores and PMA. I am able to detect IL4 and IFN-g in PBMC which I use as a positive controll for stimulation (though the conc of stimulants may not be the same for cell lines- that is the only option I have), but have not been able to get any signal from Jurkat cells. I have also tried increasing the amount of Ionomycin and PMA but to no avail. Thanks and regards to all Rana Nagarkatti Center for Biochemical Technology Molecular Immunology Division Mall Road, Delhi, India 91-110007 ____________________________________________________________________ Get free email and a permanent address at http://www.netaddress.com/?N=1
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