Electroblotting of individual polypeptides from SDS/polyacrylamide gels for direct sequence analysis
- 1 November 1987
- journal article
- research article
- Published by Wiley in European Journal of Biochemistry
- Vol. 169 (1), 9-12
- https://doi.org/10.1111/j.1432-1033.1987.tb13573.x
Abstract
A scheme for electroblotting of individual unstained protein bands from SDS/polyacrylamide gels and subsequent amino acid sequence analysis is described. Principal features are: detection of the polypeptide bands by visualization with KCl; electroblotting of excised gel pieces that correspond to the protein bands only; blotting onto polybrene-pretreated glass-fiber filter discs (12 mm diameter) placed in an electrophoretic concentrator. A high yield over all steps from gel application through electrophoresis, blotting, gas-phase sequencer degradation, and phenylthiohydantoin analysis is obtained with several different types of polypeptide (combined average yield over all steps 20%, spread 10-50%). Background is low and samples can be stored under vacuum for long periods after blotting.Keywords
This publication has 9 references indexed in Scilit:
- Domain structure of the glucocorticoid receptor protein.Proceedings of the National Academy of Sciences, 1987
- Processing of Bacillus subtilis succinate dehydrogenase and cytochrome &‐558 polypeptidesFEBS Letters, 1987
- Characterization of domain borders and of a naturally occurring major fragment of staphylococcal α‐toxinFEBS Letters, 1987
- Direct Sequence Analysis of Proteins after Electroblotting from SDS/Polyacrylamide Gel SeparationsPublished by Springer Nature ,1987
- Protein-blotting on Polybrene-coated glass-fiber sheets. A basis for acid hydrolysis and gas-phase sequencing of picomole quantities of protein previously separated on sodium dodecyl sulfate/polyacrylamide gelEuropean Journal of Biochemistry, 1985
- On the nature of the 5-lipoxygenase reaction in human leukocytes: enzyme purification and requirement for multiple stimulatory factors.Proceedings of the National Academy of Sciences, 1985
- Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.Proceedings of the National Academy of Sciences, 1979
- Rapid analysis of amino acid phenylthiohydantoins by high-performance liquid chromatographyAnalytical Biochemistry, 1977
- Rapid visuallzation of protein-dodecyl sulfate complexes in polyacrylamide gelsAnalytical Biochemistry, 1976