Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.
Open Access
- 1 September 1979
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences of the United States of America
- Vol. 76 (9), 4350-4354
- https://doi.org/10.1073/pnas.76.9.4350
Abstract
A method has been devised for the electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets. The method results in quantitative transfer of ribosomal proteins from gels containing urea. For sodium dodecyl sulfate gels, the original band pattern was obtained with no loss of resolution, but the transfer was not quantitative. The method allows detection of proteins by autoradiography and is simpler than conventional procedures. The immobilized proteins were detectable by immunological procedures. All additional binding capacity on the nitrocellulose was blocked with excess protein; then a specific antibody was bound and, finally, a second antibody directed against the first antibody. The second antibody was either radioactively labeled or conjugated to fluorescein or to peroxidase. The specific protein was then detected by either autoradiography, under UV light, or by the peroxidase reaction product, respectively. In the latter case, as little as 100 pg of protein was clearly detectable. It is anticipated that the procedure will be applicable to analysis of a wide variety of proteins with specific reactions or ligands.Keywords
This publication has 26 references indexed in Scilit:
- Detection of specific hybridoma clones by replica immunoadsorption of their secreted antibodies.Proceedings of the National Academy of Sciences of the United States of America, 1979
- Antibodies to Ribosomes in Chronic Active HepatitisGastroenterology, 1979
- Proposed uniform nomenclature for mammalian ribosomal proteinsMolecular Genetics and Genomics, 1979
- Antibodies reacting with ribosomal ribonucleoprotein in connective tissue diseasesArthritis & Rheumatism, 1979
- Immunological screening method to detect specific translation products.Proceedings of the National Academy of Sciences of the United States of America, 1978
- Evolutionary microdivergence of chick and rat liver ribosomal proteins.Online Journal of Public Health Informatics, 1977
- Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis.Online Journal of Public Health Informatics, 1977
- Improved electrophoretic and immunochemical techniques for the identification and characterization of mutant proteins, applied to ribosomal protein S8 in Escherichia coli mutantsMolecular Genetics and Genomics, 1977
- Antigen-antibody crossed electrophoresisAnalytical Biochemistry, 1965
- Méthode immuno-électrophorétique d'analyse de mélanges de substances antigéniquesBiochimica et Biophysica Acta, 1955