Description
PMA treatment of THP-1 monocytic leukemia cells is a common strategy for studies of mature monocyte function in vitro. Confluent cultures of THP-1 monocytic cells were either left untreated or treated with PMA (1uM) for 72 hrs at 37 degree C. Differentiated cells become adherent monocyte-like cells that exhibit many characteristics of mature monocytes. After 72 hrs, the cells were lysed in 1% SDS, 1.0mM sodium ortho-vanadate, 1mM sodium fluoride in 10mM Tris (pH7.4) buffer. Protein concentration was determined using the BCA method (Pierce) before diluting to final concentration and buffer