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Identification of phosphatidate phosphohydrolase purified from rat liver membranes on SDS‐polyacrylamide gel electrophoresis
Author(s) -
Siess E.A.,
Hofstetter M.M.
Publication year - 1996
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(96)00111-1
Subject(s) - polyacrylamide gel electrophoresis , membrane , chemistry , biochemistry , chromatography , centrifugation , polyacrylamide , gel electrophoresis , enzyme , microbiology and biotechnology , biology , polymer chemistry
Phosphatidate phosphohydrolase (PAP; EC 3.1.3.4) insensitive to N ‐ethyleimide was partially purified from rat liver membranes by a combination of chromatographic methods, immunoabsorption and glycerol gradient centrifugation. The specific activity was increased more than 600‐fold over that of the membrane extract. Enzyme antibodies precipitating more than 80% of PAP were obtained and used for the identification of PAP protein on SDS‐polyacrylamide gels employing the immunodetection method of Muilerman et al. [(1982) Anal. Biochem. 120, 46–51]. By this approach PAP was localized as a 31 kDa polypeptide.