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04719956001

Roche

cOmplete Lysis-M

storage temp.:room temp

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Roche

storage temp.

room temp

General description

The cOmplete Lysis-M mammalian cell protein extraction reagent contains a mild ready-to-use detergent (in 25mM bicine buffer, pH 7.6) that efficiently lyses mammalian cells and eliminates the need for scraping, sonication, or freeze-thaw cycles. It is required for the gentle extraction of proteins from both the cytoplasm and the nucleus of cultured mammalian cells. Protein yields obtained with this kit are 20 to 25% higher compared to three cycles of freeze-thaw, and approximately 20% higher than 2 minutes of sonication (with 50% pulse). Extracted proteins can be further purified or analyzed in downstream applications. Efficient lysis of mammalian cells occurs in only 5 minutes at room temperature.
The ready-to-use cOmplete Mini Protease Inhibitor Cocktail Tablets, included in the kit, function optimally with the kit′s Lysis-M reagent. The cOmplete Mini Protease Inhibitor Tablets allow the inhibition of a broad spectrum of serine, cysteine, and metalloproteases, as well as calpains. cOmplete Lysis-M reagent helps in protein extraction and simultaneously inhibit protease activity during the initial extraction steps. cOmplete Mini Protease Inhibitor Tablets contain both irreversible and reversible protease inhibitors.

Application

Lysis-M reagent is compatible with many different applications:
  • 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)
  • 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 tablets (included in the kit) contain EDTA (ethylenediaminetetraacetic acid). If the protein of interest is to be purified by IMAC (immobilized metal-chelate affinity chromatography), e.g., Poly-His tagged recombinant proteins, EDTA has to be removed (e.g., by dialysis) prior to the chromatography. Alternatively, the cOmplete Lysis-M EDTA-free product can be used.

Features and Benefits

  • 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.

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.

Preparation Note

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, metalloproteases.
Note: Each cOmplete, Mini tablet is concentrated enough to inhibit proteases in 10 ml of extract.

Other Notes

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

Legal Information

cOmplete is a trademark of Roche

Kit Components Only

Product No.
Description

  • Lysis-M Reagent for protein extraction from mammalian cells

  • cOmplete Mini Protease Inhibitor Cocktail Tablets supplied in EASYpacks

Pictograms

Exclamation mark

Signal Word

Warning

Hazard Statements

Hazard Classifications

Eye Irrit. 2 - Skin Irrit. 2

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 2

Flash Point(F)

does not flash

Flash Point(C)

does not flash


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Bryan M Ham et al.
Journal of proteome research, 7(6), 2215-2221 (2008-04-17)
Ongoing optimization of proteomic methodologies seeks to improve both the coverage and confidence of protein identifications. The optimization of sample preparation, inclusion of technical replicates (repeated instrumental analysis of the same sample), and biological replicates (multiple individual samples) are crucial

Articles

cOmplete™ Lysis-M Protocol & Troubleshooting

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