The Thermo Scientific Y-PER Yeast Protein Extraction Reagent is the first commercially available yeast lysis reagent to use a mild detergent lysis procedure to efficiently release functionally active solubilized proteins.
This detergent-based cell lysis buffer eliminates the need to use glass beads or mechanical disruption to break through the thick proteinaceous cell envelope to extract protein Y-PER Reagent is effective for S. cerevisiae and other popular species, making it applicable for use in fusion-tagged protein purification and reporter enzyme assays with these model organisms. The lysis reagent also can be used for genomic and plasmid DNA extraction from yeast.
- More protein – extracts more than twice as much protein as glass bead methods
- Less hassle – eliminates the difficulties associated with traditional glass bead lysis (e.g., clinging static-charged beads, protein-bead clumps and runaway beads)
- Optmized for yeast – validated for use with Saccharomyces cerevisiae, Schizosaccharomyces pombe and Pichia pastoris
- Versatile – effective for many different organisms, including Bacillus subtilus and both Gram-positive and Gram-negative bacteria; suitable for use in a diverse range of situations
The eukaryotic model systems have repeatedly proven their fundamental worth in studies of functional genomics and drug discovery. The unicellular nature of yeast, combined with its ability to perform post-translational modifications that closely mimic processes in higher eukaryotes, have made them important research tools. The many vectors available, as well as the development of recombinant expression systems, have given yeast an irreplaceable role in the research sector.
Yeast protein extraction and purification has traditionally been difficult and time consuming. The yeast cell is difficult to lyse because of a very complex proteinaceous cell wall that provides rigidity to the weak plasma membrane. Techniques for yeast cell lysis often involve harsh mechanical treatment while using strong reducing agents, chemicals and pH and temperature extremes. The popular glass bead lysis protocol requires special equipment and must be performed at 4°C. The low yields of protein commonly obtained with this technique are the result of denaturation and proteins nonspecifically binding to the glass beads. However, Y-PER Reagent uses a simple room temperature protocol that can be completed in 20 minutes and requires no special equipment.
The standard protocol for protein extraction with Y-PER Reagent is easy: simply add an appropriate volume of Y-PER Reagent to pelleted yeast cells, incubate at room temperature for approximately 20 minutes and spin down the debris. The resulting supernatant is a concentrated protein solution, surpassing typical yields obtained with traditional glass bead disruption.
|Thermo Scientific Y-PER Yeast Protein Extraction Reagent. Y-PER Reagent extraction of protein from two different strains each of S. cerevisiae, S. pombe, B. subtilis and E. coli. The samples were separated by 4-20% SDS-PAGE and stained with Thermo Scientific GelCode Blue Stain Reagent (Part No. 24590).
- Company, et al. (1988). Mol. Cell. Biol. 8:545.
- Jackson, J.C. and Lopes, J.M. (1996). Nucleic Acids Research 24:1322.
- Jackson, J.C. and Lopes, J.M. (1995). Gene 154:109.
- Liermann, R.T., et al. (2000). BioTechniques 28:64-65.
- Lopes, et al. (1991). Nucleic Acids Res. 19:1687.
Y-PER Plus Dialyzable Yeast Protein Extraction Reagent
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