Purification and characterization of lipopolysaccharide induced TNF-like factor from rabbit serum.
A TNF-like factor was purified from lipopolysaccharide (LPS) induced New Zealand white rabbit serum. The TNF-like factor was purified by DEAE-Sephacel, Sephacryl S-200, Mono-Q, CM-affi gel Blue, Superose 12 H/R preparative columns to the specific activity of 4 x 10(6) U/mg protein. The purified protein was 45 kDa in its oligomeric form and 22 kDa in its monomeric form. Rabbit TNF-like factor had a pI value of 5.0 and was resistant to trypsin digestion. The TNF-like factor reacted with polyclonal-Ab against human TNFalpha on immunoblot and immunoprecipitation analysis and interacted with human TNF receptors. Taken together, rabbit TNF-like factor might be a high molecular weight form of rabbit TNFalpha.
Lee
T H
TH
Department of Biochemistry, College of Science and Bioproducts Research Center, Yonsei University, Seoul, South Korea.
Rhim
T Y
TY
Son
O K
OK
Kim
Y H
YH
Kim
S S
SS
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001667:Binding, Competitive; D003001:Cloning, Molecular; D006801:Humans; D007525:Isoelectric Focusing; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008970:Molecular Weight; D011817:Rabbits; D018124:Receptors, Tumor Necrosis Factor / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis* / Q000737:chemistry / Q000276:immunology
D000911:Antibodies, Monoclonal; D008070:Lipopolysaccharides; D018124:Receptors, Tumor Necrosis Factor; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha