Rapid protein extraction from Saccharomyces cerevisiae
- 1 October 1994
- Vol. 10 (10) , 1305-1310
- https://doi.org/10.1002/yea.320101007
Abstract
We have developed and evaluated an easy and rapid method for extraction of proteins from yeast cells for sodium dodecyl sulphate (SDS)–gel electrophoresis and Western blotting. The procedure comprises a centrifugation step to harvest the cells, addition of a sample buffer and heating, then another centrifugation step before applying the extracted proteins found in the supernatant to an SDS gel. It is applicable to the study of large numbers of samples in 1 day. This procedure is easier, quicker, and as efficient as procedures using base and 2‐mercaptoethanol, but somewhat less efficient than lysis with glass beads under certain conditions.Keywords
This publication has 14 references indexed in Scilit:
- Analysis of the sequence requirements for glycosylphosphatidylinositol anchoring of Saccharomyces cerevisiae Gas1 proteinJournal of Biological Chemistry, 1993
- end3 and end4: two mutants defective in receptor-mediated and fluid-phase endocytosis in Saccharomyces cerevisiae.The Journal of cell biology, 1993
- Determinants for glycophospholipid anchoring of the Saccharomyces cerevisiae GAS1 protein to the plasma membrane.Molecular and Cellular Biology, 1991
- [48] Yeast endocytosis assaysPublished by Elsevier ,1991
- [45] Analysis of polypeptide transit through yeast secretory pathwayPublished by Elsevier ,1991
- [35] Epitope tagging and protein surveillancePublished by Elsevier ,1991
- Extraction and rapid inactivation of proteins from Saccharomyces cerevisiae by trichloroacetic acid precipitationYeast, 1989
- Two nuclear mutations that block mitochondrial protein import in yeast.Proceedings of the National Academy of Sciences, 1984
- Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.Proceedings of the National Academy of Sciences, 1979
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970