Jose: You can try MTT, a water soluble tetrazolium salt, which is a yellow solution that is converted to an insoluble purple formazan when cleaved by dehydrogenase enzymes. The insoluble formazan can be solubilized isopropanol and then measured on a spectrophotometer. Sigma sells MTT (Product # M5655) relatively cheap. In my hands it worked really well for mitogens but was a bit less sensitive for recall antigens (we used it at 5mg/ml). Some have used CD69 and flow for this also, correlations were good, however, you'll have to deal with the issue that cellular activation doesn't always lead to cellular proliferation. Joanne -----Original Message----- From: Jose Benito <jbenito1@hotmail.com> To: cyto-inbox Date: Tuesday, September 19, 2000 6:57 PM Subject: proliferative responses > >Hi everyone, I am trying to set up a colorimetric assay to measure >lymphoproliferative responses to mitogens and recall antigens. I have tried >the Alamar Blue without any sucess. I will probably try also the celltiter >assay from promega. Does anyone have experience with this kind of assays? >Can they really substitute the 3Hthymidine assay? >Thanks >Jose Miguel >Hospital Carlos III >Madrid >Spain >_________________________________________________________________________ >Get Your Private, Free E-mail from MSN Hotmail at http://www.hotmail.com. > >Share information about yourself, create your own public profile at >http://profiles.msn.com. >
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