Thermo Scientific Y-PER-Plus Dialyzable Yeast Protein Extraction Reagent effectively lyses yeast and bacterial cells to extract and solubilize proteins for analysis.
This proprietary formulation of cell lysis buffer facilitates protein extraction from S. cerevisiae and S. pastoris yeast cell cultures. Glass bed disruption is not necessary; protein extraction is achieved by a 20-minute incubation in the reagent at room temperature. Unlike other cell lysis buffers, the Y-PER-Plus Reagent uses a detergent component that is fully dialyzable, allowing complete exchange of the extracted protein into any buffer required by downstream techniques.
- Stable formulation designed for room temperature storage
- Yields protein extract with simple 20-minute hands-off incubation
- Requires no rigorous mechanical disruption or lengthy enzymatic treatments
- Tris-based buffer has a very low ionic strength and does not include sodium chloride
- Simple formulation contains no reducing agents or chelating agents
- Fully dialyzable formulation provides complete control over downstream applications
- Compatible with both the Thermo Scientific Pierce BCA and Coomassie Plus Protein Assays
Product Details (Recommendations for Use):
- Fresh Cells and Frozen Cells: The Y-PER-Plus Reagent is capable of extracting proteins equally well from both freshly harvested and frozen cells
- Cell Density and Strain Variation: Differences in growth rate among organisms, growth temperature and media composition affects the number of cells harvested from a given culture volume. Therefore several suggestions for the amount of Y-PER-Plus Reagent to use for a given cell pellet (wet cell paste) weight are included.
- Pichia pastoris: To achieve lysis of cells grown in rich media such as YEPD, cells must be harvested during log-phase growth. To enhance lysis of cells in stationary phase, add DTT or TCEP-HCl (to a final concentration of 0.1M and 20 to 50mM, respectively) directly to the Y-PER-Plus Reagent before lysis to release much more of the soluble proteins. For best results use 2.5 to 5.0mL of Y-PER-Plus Reagent per 1 gram of cell paste; scale up or down accordingly. An alternative to adding reducing agents is adding a protease inhibitor and incubating at 45°C.
- Saccharomyces cerevisiae: For best results, harvest cells in log-phase. To increase yield for cells in stationary-phase, add DTT or TCEP•HCl (to a final concentration of 0.1M and 20 to 50mM, respectively) directly to the Y-PER-Plus Reagent before extraction. No differences in efficiency results for either rich- or synthetic-defined media. For best results use 2.5 to 5.0mL of Y-PER Plus Reagent per 1 gram of cell paste; scale up or down accordingly.
- Schizosaccharomyces pombe: For best results, use the original Y-PER Reagent (Part No. 78990) for extracting proteins from S. pombe.
- Bacillus subtilis: The Y-PER-Plus Reagent will not extract protein from B. subtilis spores. When using a strain that is able to sporulate, harvest cells during log-phase. For strains unable to sporulate, cells may be grown to saturation before harvesting. Use 2.5 to 5.0mL of Y-PER-Plus Reagent per 1g of cell paste; scale up or down accordingly.
- Escherichia coli: Use 2.5 to 5.0mL of Y-PER-Plus Reagent per 1g of cell paste; scale up or down accordingly.
- Enzyme Activity: Because all proteins differ in structure, solubility and stability, a particular protein might not retain optimum activity in the presence of Y-PER-Plus Reagent. For example, the original Y-PER Reagent works well for assaying beta-galactosidase activity, but Y-PER-Plus Reagent slows down its kinetics. Y-PER-Plus Reagent is compatible with standard affinity-purification protocols for glutathione S-transferase (GST) and His-tagged proteins.
Y-PER Yeast Protein Extraction Reagent
B-PER Bacterial Protein Extraction Reagents