Purification of an autocrine growth factor in conditioned medium obtained from primary cultures of scleral fibroblasts of the chick embryo.
Scleral fibroblasts of the chick embryo were found to secrete autocrine growth factors. One of the factors was purified from conditioned medium collected from growing-phase cultures of these cells by DEAE-Sepharose column chromatography and following non-denaturing polyacrylamide gel electrophoresis. The specific activity was increased 1100-fold by this purification. The chromatographically purified growth factor was still active after incubation at 95 degrees C, at pH 10 or pH 3, or with glycosidase H, but inactive after incubation with dithiothreitol or trypsin. An active protein having a molecular weight of 32 kDa was found to be the major component of the final preparation.
Fujioka
M
M
Faculty of Integrated Arts and Sciences, Hiroshima University, Japan.
Shimamoto
N
N
Kawahara
A
A
Amano
M
M
Watanabe
K
K
D000818:Animals; D002356:Cartilage / Q000166:cytology; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D002642:Chick Embryo; D002847:Chromatography, Agarose; D003470:Culture Media; D004229:Dithiothreitol / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism*; D006031:Glycosylation; D006133:Growth Substances / Q000032:analysis / Q000302:isolation & purification* / Q000494:pharmacology; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration
D003470:Culture Media; D006133:Growth Substances; D004229:Dithiothreitol