We have found MRK16 to give the highest level of fluorescence of all the commercially available anti-P-glycoprotein mAbs that we have tried. It is available from Kamiya (Kamiyabiomedical.com) only as a purified antibody and we use a goat anti mouse IgG2a PE as a second step reagent. A good control is normal human peripheral blood. After staining with the anti-Pgp antibody, counterstain with mAbs to CD4, CD8, and an NK marker. NK cells have the highest level of Pgp, followed by CD8 cells and then CD4 cells. The monocytes and granulocytes in your sample will be negative for Pgp. These lymphocytes all have much lower levels of Pgp than do resistant cell lines produced in vitro, but they reflect what you will see in patient samples. Barbara J Taylor Flow Cytometry Research Lab Children's National Medical Center Washington DC btaylor@cnmc.org
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