Partial purification of murine B cell stimulatory factor (BSF)-1.
BSF-1 was partially purified from serum-free culture supernatants of cells of the EL-4 thymoma line, which had been induced 48 hr earlier with 4 beta-phorbol-12 beta-myristate-12 alpha-acetate (PMA). BSF-1 in 10-liter batches was adsorbed onto and eluted from trimethylsilyl-controlled pore glass beads (TMS-CpG) and then subjected to reverse-phase high-performance liquid chromatography (RP-HPLC). The recovery of BSF-1 activity by TMS-CpG and RP-HPLC ranged from 52 to 55% and 187 to 227%, respectively. The specific activity in units per milligram of protein of partially purified BSF-1 was approximately 2600 times higher than that of the culture supernatant protein. The partially purified BSF-1 had a single isoelectric point of 6.3 and an apparent m.w. between 18,000 and 21,700 when analyzed by isoelectric focusing and gel filtration-HPLC, respectively. The ability to prepare large amounts of partially purified BSF-1 by a rapid and efficient procedure should be of great help in both biochemical and immunologic studies of this lymphokine.
D000818:Animals; D002460:Cell Line; D002474:Cell-Free System; D005591:Chemical Fractionation / Q000379:methods; D002851:Chromatography, High Pressure Liquid; D003470:Culture Media / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D006133:Growth Substances / Q000032:analysis / Q000302:isolation & purification*; D015847:Interleukin-4; D008222:Lymphokines / Q000032:analysis / Q000302:isolation & purification*; D051379:Mice; D012967:Sodium Dodecyl Sulfate; D013945:Thymoma / Q000032:analysis / Q000276:immunology; D014297:Trimethylsilyl Compounds
D003470:Culture Media; D006133:Growth Substances; D008222:Lymphokines; D014297:Trimethylsilyl Compounds; D015847:Interleukin-4; D012967:Sodium Dodecyl Sulfate