Figure 2. Display of several phospho-specificities.
1 X106 serum starved (12 hrs) U937 cells were stimulated with either anisomycin (20 mM), PMA/IO (500 ng/ml), PHA (50 mg/ml), IFNg (200 ng/ml), IL-4 (200 ng/ml), or GMCSF (200 ng/ml) for 15 min. Cells were fixed, permeabilized and stained (as per protocol 1) with phospho-p38(T180/Y182)-AX647 (clone 36), phospho-p44/42(T202/Y204)-AX647 (clone 20a), PY20-PE, phospho-STAT1(Y701)-AX488 (clone 14), phospho-STAT6(Y641)-AX488 (clone 18), phospho-STAT5(Y694)-AX488 (clone 47). Antibodies were used at 0.125 mg.