Premium
Production of monoclonal antibodies specific to the carboxyl terminal region of the 85 kDa subunit of phosphatidylinositol 3‐kinase: Use of the antibodies in recognition of mutant p85
Author(s) -
DADUANG SAKDA,
NAGATA SATOSHI,
MATSUDA MICHIYUKI,
YAMORI TAKAO,
ONODERA KAZUKIYO,
FUKUI YASUHISA
Publication year - 1995
Publication title -
immunology and cell biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.999
H-Index - 104
eISSN - 1440-1711
pISSN - 0818-9641
DOI - 10.1038/icb.1995.21
Subject(s) - monoclonal antibody , epitope , mutant , microbiology and biotechnology , protein subunit , phosphatidylinositol , immunoprecipitation , antibody , proto oncogene tyrosine protein kinase src , kinase , biology , chemistry , biochemistry , gene , genetics
Summary We have established two hybridomas producing mAb to the carboxyl terminal region of phosphatidylinositol‐3 kinase 85 kDa subunit type α (p85α). Analysis using deletion mutants of p85 revealed that epitopes for the two mAb were located on the border of the src homology 2 (SH2) sequence located at the carboxyl end of p85. They immunoprecipitated free p85 efficiently, but reactivity to p85 bound to pi 10 was very weak. Together with the mAb which we have reported previously, a panel of mAb that covered the various parts of p85α was obtained. Using this panel, we characterized two mutants of p85 (70 and 50 kDa) expressed in the human colon carcinoma cell line, HCC2998. No wild‐type p85 was detected in these cells. A mAb specific to the carboxyl terminal region detected p70 but not p50, suggesting that this region is missing in p50. The panel of mAb is a useful tool to use to analyse mutant forms of p85.