title: Characterization of macrophage colony-stimulating factor in body fluids by immunoblot analysis.
abstract: We characterized the molecular species of human macrophage colony-stimulating factor (hM-CSF) found in serum and urine, using immunoblot analysis after partial purification on an antibody-bound affinity column. Although antibodies were prepared using the recombinant product of the large form of hM-CSF with a molecular weight (MW) of 85 Kd as the antigen, this immunoblot system was also capable of detecting the small form of hM-CSF with a MW of 40 to 60 Kd. A single band with a MW of 43 Kd, which reacted with anti-recombinant hM-CSF IgG but not with control IgG, was found when serum and urine from normal adults underwent electrophoresis on reduced sodium dodecyl sulfate-polyacrylamide gel and subsequent immunoblotting. This band represented a subunit of the large form of hM-CSF, because the large form of hM-CSF is a homodimer of a subunit with a MW of 43 Kd and the small form of hM-CSF is a homodimer of a subunit with a MW of 20 to 30 Kd. Analysis of serum and urine from leukemic patients and pregnant women, who had higher serum levels of hM-CSF than normal adults, showed only a single band with a MW of 43 Kd as a hM-CSF-specific molecule. These results suggest that the large form of hM-CSF is the major species in human body fluids.
authors.affiliation: Biochemical Research Laboratory, Morinaga Milk Industry Co, Ltd, Kanagawa, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Suzu
authors.forename: N
authors.initials: N
authors.lastname: Yanai
authors.forename: Y
authors.initials: Y
authors.lastname: Sato-Somoto
authors.forename: M
authors.initials: M
authors.lastname: Yamada
authors.forename: T
authors.initials: T
authors.lastname: Kawashima
authors.forename: T
authors.initials: T
authors.lastname: Hanamura
authors.forename: N
authors.initials: N
authors.lastname: Nagata
authors.forename: F
authors.initials: F
authors.lastname: Takaku
authors.forename: K
authors.initials: K
authors.lastname: Motoyoshi
mesh_terms: D001826:Body Fluids / Q000737:chemistry*; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D015151:Immunoblotting; D007074:Immunoglobulin G / Q000032:analysis; D007938:Leukemia / Q000097:blood / Q000652:urine; D016173:Macrophage Colony-Stimulating Factor / Q000032:analysis* / Q000737:chemistry; D008970:Molecular Weight; D011247:Pregnancy
chemical_list: D007074:Immunoglobulin G; D016173:Macrophage Colony-Stimulating Factor