An efficient, ready-to-use mammalian cell lysis solution that eliminates scraping, freeze-thaw and sonication methods.

Thermo Scientific M-PER Mammalian Protein Extraction Reagent is designed to provide highly efficient total protein extraction from cultured mammalian cells. The complete cell lysis reagent is a nondenaturing detergent formulation that dissolves cell membranes and extracts total cellular protein in only 5 minutes. M-PER Reagent requires little or no mechanical disruption and yields more protein than freeze/thaw cycles and sonication. This mammalian cell lysis reagent is so efficient that adherent cells do not need to be scraped from the culture dish, enabling easy and direct lysis and analysis of cells grown in 24-well and 96-well plates. Resulting cell lysates from either adherent and suspension cells are compatible with many downstream assays including immunoassays, enzyme assays and a variety of common reporter assays.

Highlights:

• Gentle – mild detergent lysis, yielding extracts that are immediately compatible with coomassie (Bradford) and BCA protein assays or SDS-PAGE
• Compatible – extracts soluble proteins in nondenatured state, enabling direct use in immunoprecipitation and other affinity purification procedures
• Amine-free and dialyzable – formulation ensures compatibility with subsequent assay systems
• Convenient – lyse adherent cells directly in plate or after scraping and washing in suspension
• Non-denaturing – maintain activity of luciferase, beta-galactosidase, CAT and other reporter genes as well or better than other suppliers' products and freeze/thaw methods

Product Details:

Protein extraction is usually the first key step in any proteomics analysis procedure. To extract protein from cultured cells it is necessary to perform cell lysis, thus opening the cell and releasing the proteins of interest. Several methods are commonly used to physically lyse cells, including mechanical disruption, liquid homogenization, sonication, freeze/thaw cycles and manual grinding.

M-PER Reagent compatibility with reporter assays in transiently transfected mammalian cells. FM2 cells were transiently transfected with a reporter construct containing the luciferase gene. The transfected cells were lysed with either M-PER Reagent or Brand P lysis buffer and subjected to luciferase assay. For β-galactosidase and CAT assays, MDA-MB-231 cells were contransfected with reporter constructs expressing β-Gal and CAT, respectively. The transfected cells were lysed with M-PER Reagent or the freeze-thaw method, and the lysates were assayed for activity.

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