Dear experts in flow cytometry, I am performing an intracellular staining of lymphocytes for INF and TNF. These markers work very well after PMA or peptide stimulation. My problem is that all INF or TNF positives cells are CD69 neg. I have tried CD69 from Immutech and from BD. Both works well on non-fixed cells. Unfortunately, I cannot visualize CD69+ after fixation. I have try to fix with perm&fix kit (Caltag), BD fixation sol. and classical saponine/formaldehyde method. I have also tried to stain with CD69 before and after fixation. I have always had all CD69 at 101log (same for PMA and peptide activation and non activated control cells). Can someone tell me what I am doing wrong, please ? Thank you for all your help and any ideas ! Michal -- Michal Abel, MD, PhD Laboratory of Clinical Immunology INSERM U25 Necker Hospital, 161 Rue de Sèvres. Paris tel: 33 1 42 19 28 87 fax: 33 1 44 49 53 74
This archive was generated by hypermail 2b29 : Sun Jan 05 2003 - 19:26:20 EST