Murine megakaryocyte colony stimulating factor: its relationship to interleukin 3

A protocol for the partial purification of murine megakaryocyte colony stimulating factor from WEHI-3 cell conditioned medium is described. The procedure involving separation by anion exchange chromatography and molecular sieving on Sephadex G100, gave a 260-fold purification over starting material, as determined by in-vitro megakaryocyte colony formation. Fifty percent of maximum colony numbers were obtained at a dose of 1 microgram protein/ml. Electrophoretic analysis of the partially purified preparation followed by silver nitrate staining revealed four major protein bands with apparent molecular weights of 67 Kd, 43 Kd, 38 Kd and 28 Kd. All megakaryocyte colony stimulating activity and interleukin 3 activity was found in the 22-25 Kd fraction from a preparative SDS-polyacrylamide gel using non-reducing conditions, a molecular weight range that coincided with the 28-Kd molecule obtained when reducing conditions were employed.