Y-PER Plus Dialyzable Yeast Protein Extraction Reagent

Description

Thermo Scientific Y-PER-Plus Dialyzable Yeast Protein Extraction Reagent effectively lyses yeast and bacterial cells to extract and solubilize proteins for analysis.

Features of Y-PER Plus Dialyzable Yeast Protein Extraction Reagent:

• 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 , allowing complete exchange of the extracted protein into any buffer required by downstream techniques
• 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 50 mM, 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 5.0 mL 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 50 mM, 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 5.0 mL 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 5.0 mL of Y-PER-Plus Reagent per 1g of cell paste; scale up or down accordingly.

Escherichia coli: Use 2.5 to 5.0 mL 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.

Specifications

Reagent Type: Cell Lysis Buffer, Detergent Solution
Form: Liquid
Product Line: Y-PER™
Product Size: 500 mL

Contents & storage

Store product at room temperature.

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