We use immobilized anti-CD3 mAb (coated on culture wells, at 4'C overnight in PBS) to stimulate bovine lymphocytes and have success in staining for intracellular IFN-gamma. (In fact large % of lymphocytes produce IFN-gamma when stimulated by immobilized anti-CD3 mAb, compared to ConA or PMA/Ionomycin stimulation). Hope would be the same for human lymphocytes. We have not tried soluble mAb. You may try both immobilized and soluble mAb for the first time abd then select one that works. Good luck Sathi
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