COMPLETE LYSIS MAMMALIAN EDTA-FREE

General description

The cOmplete^® Lysis-M EDTA-free mammalian cell protein extraction reagent contains a mild detergent (in 25mM bicine buffer, pH 7.6). cOmplete Lysis-M EDTA-free is intended for the efficient and gentle extraction of proteins from both the cytoplasm and the nucleus of cultured mammalian cells. Efficient lysis of mammalian cells occurs in only 5 minutes at room temperature, eliminating the need for scraping, sonication, or freeze-thaw cycles.
The ready-to-use cOmplete Mini EDTA-free Protease Inhibitor Cocktail Tablets, included in the kit, work optimally with the kit ′s Lysis-M reagent. cOmplete Lysis-M EDTA-free provides a very convenient and efficient way to extract proteins and simultaneously inhibit protease activity during the initial extraction steps. cOmplete Mini EDTA-free Protease Inhibitor Cocktail Tablets contains both irreversible and reversible protease inhibitors. cOmplete Mini EDTA-free tablets efficiently inhibit a wide range of serine and cysteine proteases, but not metalloproteases.
The extracted proteins can be further purified using immobilized metal chelate affinity chromatography (IMAC) or analyzed in downstream applications.

Application

Lysis-M reagent is compatible with many different applications, including reporter assays (e.g., β-galactosidase, luciferase, chloramphenicol acetyltransferase), immunoassays (e.g., Western blots, ELISAs, RIAs), protein assays (e.g., protein kinase A, protein kinase C, and tyrosine kinase), and protein purification. Furthermore, the cell lysate is compatible with protein assays such as Coomassie staining and BCA (2′-benzoyloxycinnamaldehyde) protein assays, and the reagent can be removed by dialysis.
Note: cOmplete Mini EDTA-free tablets (included in the kit) are employed to stabilize those extracts where the activity of metal-containing proteins must not be affected. Since EDTA interferes with IMAC (immobilized metal-chelate affinity chromatography), cOmplete Mini EDTA-free is preferentially used in the isolation process of Poly-His tagged fusion proteins or subsequent assays.

Features and Benefits

• Compltely and gently lyses mammalian cells in just 5 minutes.
• Saves time by eliminating scraping, sonication, and freeze-thaw cycles.
• Produce higher yields of extracted protein than sonication.
• Preserves protein structure, since reagents will not denature or otherwise alter proteins.
• Saves money, since one product protects against many proteases.
• Saves steps, since extracted proteins may be used directly in many downstream procedures (even those that would be affected by EDTA, such as IMAC).
• Intended use: Simultaneous extraction and protection of soluble proteins from mammalian cells.
• Sample material: Mammalian cells in culture (e.g., Cos-7 cells, harvested at confluency).
• Time required: 5 minutes (cell lysis at room temperature).
• Number of extractions: 20 (10 ml each).
• Effective against: Serine proteases, cysteine proteases (but not metalloproteases).

Packaging

1 kit containing 2 components. Each tablet is sufficient for 10 mL of extract. For the lysis of up to 100 g of mammalian cells.

Storage and Stability

Store cOmplete Protease Inhibitor Tablets at 2–8 °C, Lysis-M-reagent at 15–25 °C.

Other Notes

For life science research only. Not for use in diagnostic procedures.

Legal Information

Roche is a registered trademark of Roche Molecular Systems, Inc.

cOmplete is a trademark of Roche