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Protocol for Whole Cell Lysis of Yeast
Published: Jan 5, 2011 DOI: 10.21769/BioProtoc.14 Views: 43017
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Abstract
This protocol describes how to perform lysis on whole yeast cell samples using NaOH. The lysed cells can then be used for downstream applications such as the extraction of total proteins.
Materials and Reagents
- NaOH (Sigma-Aldrich)
- Beta-mercaptoethanol (beta-ME) (Sigma-Aldrich)
- 100% (w/v) trichloroacetic acid (TCA) (Sigma-Aldrich)
- Pepstatin A (Sigma-Aldrich, catalog number: P5318 )
- Leupeptin (Sigma-Aldrich, catalog number: L2884 )
- PMSF (Sigma-Aldrich, catalog number: P7626 )
- Acetone (Sigma-Aldrich)
- Protease inhibitors
- DMSO
- Glycerol
- BPB
- Ethanol
- SDS lysis buffer
- 7.5 ml 1 M Tris-HCl (pH 6.8) (see Recipes)
- DTT stock (see Recipes)
- 5x SDS-PAGE gel loading dye (see Recipes)
- Protease inhibitors (see Recipes)
Procedure
Copyright: © 2011 The Authors; exclusive licensee Bio-protocol LLC.
Category
Biochemistry > Protein > Electrophoresis
Cell Biology > Cell viability > Cell lysis
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