Hello Unni, the difference is to my knowledge that immobilized anti-CD3 mAb should also work with purified T cells, on the other hand soluble mAb gives you T cell proliferation and cytokine production just in the presence of adherent monocytes (the mAb is bound to their Fc receptors and immobilized/and concentrated that way, in my hands it works well down to 4 ng/ml, of course it is dependent on monocyte numbers). So, I would suggest the immobilized assay (I have used 10ug/ml of purified mAb), this should be better for costimulation using anti-CD28 mAb and also allows you to work with purified T cells or lymphocytes, that gives you clean background if you get rid of monocytes. Also bear in mind that adherent monocytes are strongly activated and could affect the cytokine milieu in your cultures. -- Karel Drbal Laboratory of Leukocyte Antigens Institute of Molecular Genetics Academy of Sciences of the Czech Republic Videnska 1083 142 20 PRAHA 4 Czech Republic, Europe voice: +420-2-4752589 fax: +420-2-44472282 home: +420-2-360016 e-mail: drbal@leuko.biomed.cas.cz WWW: http://leuko.biomed.cas.cz Unni Haddeland wrote: > > Is soluble or immobilized CD3mab preferable for lymphocyte activation? > > I will try to activate lymphocytes by CD3 mab (together with soluble CD28 mab > and possibly after antigen stimulation) and measure intracellular cytokines by > Flow. In some papers soluble anti-CD3 (conc. 4 microgram/ml) is used, but in > most papers immobilized anti CD3 is used. Is there any difference between the > two? > Unni Haddeland, ph.D. > Research Laboratory > University College of Akershus > Ringstabekkvn 105 > 1365 Bekkestua > NORWAY
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