Hello, everyone, It seems possible to me that cytokine receptors may be detected by flow without using anti-receptor Mabs or biotinylated ligands. The idea is to incubate target cells with saturating concentrations of unlabeled ligand, wash the cells and detect receptor-bound ligand with an appropriate Mab. Certainly, the Mab must recognize epitopes not involved in receptor-ligand interaction. Same approach is used in cytokine ELISA, where a "capturing" antibody (cytokine receptor in the above example) targets one epitope, while "developing" antibody targets the other. We would like to check the presence of IFN receptors on some tumor cell lines but don't like the idea to spend over $300 on anti-receptor Mab to run just a couple of tests. From the other hand, we are interested in buying some anti-IFN-gamma Mabs to study Th1 biased responses. I understand that policlonal anti-IFN antibodies should do the trick, but I am not sure about level of non-specific binding during indirect staining. I appreciate if anyone can send me some references or personal experience on this matter (i.e., whether a given clone can bind receptor-bound interferon-gamma :-J ). Thanks in advance, Yuri YKudinov@CHW.edu ----------------------------- Yuri Kudinov, postdoctoral fellow Immunotherapy Laboratory St. Vincent Medical Center 201 South Alvarado Street Prof.Office Bldg, Ste 312 Los Angeles, CA 90057 ----------------------------- "Bad appetite does not necessarily mean holiness" Sir George Bernard Shaw (1856-1950) -----------------------------
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