Purification and biochemical characterization of human pluripotent hematopoietic colony-stimulating factor.
Pluripotent hematopoietic colony-stimulating factor (pluripotent CSF), a protein that is constitutively produced by the human bladder carcinoma cell line 5637, has been purified from low serum (0.2% fetal calf serum)-containing conditioned medium. The purification involved sequential ammonium sulfate precipitation, ion-exchange chromatography, gel filtration, and reversed-phase high-performance liquid chromatography. The purified protein has a molecular weight of 18,000 in NaDodSO4/polyacrylamide gel electrophoresis, both by the silver staining technique and by elution of biological activity from a corresponding gel slice, and has an isoelectric point of 5.5. Pluripotent CSF supports the growth of human mixed colonies, granulocyte-macrophage colonies, and early erythroid colonies and induces differentiation of the human promyelocytic leukemic cell line HL-60 and the murine myelomonocytic leukemic cell line WEHI-3B (D+). The specific activity of the purified pluripotent CSF in the granulocyte-macrophage colony assay is 1.5 X 10(8) units/mg of protein.
D001681:Biological Assay; D002460:Cell Line; D003115:Colony-Stimulating Factors / Q000302:isolation & purification*; D003470:Culture Media; D006412:Hematopoietic Stem Cells / Q000502:physiology; D006801:Humans; D007526:Isoelectric Point; D008970:Molecular Weight; D001749:Urinary Bladder Neoplasms / Q000032:analysis
D003115:Colony-Stimulating Factors; D003470:Culture Media