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Recovery of the contact site A glycoprotein of Dictyostelium discoideum from sodium dodecyl sulfatepolyacrylamide gel and characteristics of monoclonal antibodies against the recovered protein
Author(s) -
Ochiai Hiroshi,
Jin Kimiaki,
Kakihara Noriaki,
Saito Tamao,
Fukuzawa Masashi
Publication year - 1990
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150111015
Subject(s) - dictyostelium discoideum , glycoprotein , monoclonal antibody , gel electrophoresis , antibody , electroelution , polyacrylamide gel electrophoresis , biochemistry , microbiology and biotechnology , chemistry , biology , immunology , enzyme , gene
Monoclonal antibodies were produced against a cell‐cell adhesion (contact site A) glycoprotein of Dictyostelium discoideum , isolated by preparative gel electrophoresis. The glycoprotein was recovered by electroelution from a polyacrylamide gel strip and used for the production of monoclonal antibodies. Four of the five antibodies obtained bound specifically to the protein moiety of the contact site A glycoprotein. The specificities of the antibodies were in striking contrast to those of antibodies raised against the contact site A glycoprotein purified by Triton X‐114 phase separation and DEAE chromatography. The majority of the latter antibodies recognized the carbohydrate moiety of the contact site A glycoprotein and cross‐reacted heavily with other membrane glycoproteins.