PMID: 6165580Mar 1, 1981Paper

Immunological detection of specific proteins in total cell extracts by fractionation in gels and transfer to diazophenylthioether paper

European Journal of Biochemistry
J Reiser, J Wardale

Abstract

We describe a sensitive immunological procedure for the detection of specific proteins in total cell extracts and for the comparison of antigenically related polypeptides. Proteins are fractionated in polyacrylamide gels and transferred electrophoretically to diazophenylthioether paper, to which they bind covalently. Specific proteins are identified by incubation with specific antibody and 125 I-labeled protein A from Staphylococcus aureus, followed by autoradiography. High-resolution separation of proteins prior to transfer is achieved by polyacrylamide gradient gel electrophoresis in the presence of sodium dodecyl sulfate or by nonequilibrium pH gradient electrophoresis, followed by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. Further information can be obtained by limited enzymatic proteolysis of the proteins in the gel following polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate and analysis of the cleavage products by gel electrophoresis at right angles to the first gel. We show the application of this technique to the detection and comparison in extracts from infected cells of proteins related immunologically to the simian virus 40 capsid proteins VP1 and VP3.

References

Jul 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·J RenartG R Stark
Sep 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·H TowbinJ Gordon
Dec 1, 1976·Proceedings of the National Academy of Sciences of the United States of America·K Burridge
Sep 1, 1974·Proceedings of the National Academy of Sciences of the United States of America·H Heitzmann, F M Richards
May 1, 1971·Journal of Virology·M K EstesJ S Pagano
Jan 25, 1980·Nucleic Acids Research·E J Stellwag, A E Dahlberg

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Citations

Sep 4, 1984·Journal of Immunological Methods·H Towbin, J Gordon
Nov 20, 1986·Journal of Immunological Methods·Y ChemlaJ Sperling
May 12, 1989·Journal of Immunological Methods·D I Stott
May 1, 1982·Proceedings of the National Academy of Sciences of the United States of America·K KakitaM A Permutt
May 1, 1984·Proceedings of the National Academy of Sciences of the United States of America·D RoyA Spector
Jan 1, 1983·Archives of Virology·S van DawenS Roth
Jan 1, 1986·Cell Biochemistry and Function·P J Van der Sluis, G J Boer
May 1, 1983·Analytical Biochemistry·J M Gershoni, G E Palade
Apr 30, 1985·Biochemical and Biophysical Research Communications·A A Davies, M J Crumpton

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