04719948001
Roche
cOmplete™ Lysis-B (2x) EDTA-free
usage
sufficient for 20 extractions
manufacturer/tradename
Roche
shipped in
ambient
General description
The cOmplete Lysis-B (2x) EDTA-free bacterial protein extraction reagent contains a mild, double-concentrated, non-ionic detergent in 20 mM Tris/HCl (pH 7.5) that enables very efficient and gentle extraction of proteins, especially recombinant proteins, from bacteria (e.g., E. coli) in small volumes. This simple extraction method eliminates the need for mechanical disruption (e.g., standard sonication) and produces the highest possible protein yields.
The ready-to-use cOmplete Mini EDTA-free Protease Inhibitor Cocktail Tablets, included in the kit, function optimally with the kit ′s Lysis-B reagent. cOmplete Lysis-B (2x) EDTA-free inhibitor provides a very convenient and efficient way to extract proteins and simultaneously inhibit protease activity during the initial extraction steps. cOmplete Mini EDTA-free inhibitor contains both irreversible and reversible protease inhibitors.
Extracted proteins can be further purified using immobilized metal chelate affinity chromatography (IMAC) or analyzed in downstream applications.
The ready-to-use cOmplete Mini EDTA-free Protease Inhibitor Cocktail Tablets, included in the kit, function optimally with the kit ′s Lysis-B reagent. cOmplete Lysis-B (2x) EDTA-free inhibitor provides a very convenient and efficient way to extract proteins and simultaneously inhibit protease activity during the initial extraction steps. cOmplete Mini EDTA-free inhibitor contains both irreversible and reversible protease inhibitors.
Extracted proteins can be further purified using immobilized metal chelate affinity chromatography (IMAC) or analyzed in downstream applications.
Application
cOmplete™ Lysis-B (2x) EDTA-free is intended for the rapid cell lysis of bacterial cells in only 10 minutes with simultaneous inhibition of protease activity in the cell lysate. cOmplete Lysis-B (2x) EDTA-free enables highly efficient protein extraction from several common bacterial host strains (especially E. coli BL21 strains) and the simultaneous inhibition of a multitude of proteases, including serine proteases and cysteine proteases, but not metalloproteases.
The Lysis-B reagent purifies inclusion bodies to near-homogeneous levels, but does not solubilize them. The Lysis-B reagent has also been successfully used for the extraction of proteins from insect cells infected by baculovirus.
cOmplete Lysis-B (2x) EDTA-free extracts proteins from fresh and frozen cells. The extracted proteins are suitable for further purification or analysis in downstream applications. As Lysis-B Reagent is based on a Tris buffer system, Tris-HCl buffers are recommended for subsequent protein purification.
When extractions or single-step isolations are necessary in the acid pH range, simply include pepstatin along with cOmplete Mini EDTA-free tablets to ensure aspartic (acid) protease inhibition.
It has been used for cell incubation in the transient transfection assay, detection of expressed protein and western blot analysis.
Note: cOmplete Mini EDTA-free tablets (included in the kit) are employed to stabilize those protein extracts where activity of metal-containing proteins must not be affected. Since EDTA interferes with IMAC (immobilized metal affinity chromatography), cOmplete Mini EDTA-free is preferentially used in the isolation process of Poly-His tagged fusion proteins or subsequent assays.
The Lysis-B reagent purifies inclusion bodies to near-homogeneous levels, but does not solubilize them. The Lysis-B reagent has also been successfully used for the extraction of proteins from insect cells infected by baculovirus.
cOmplete Lysis-B (2x) EDTA-free extracts proteins from fresh and frozen cells. The extracted proteins are suitable for further purification or analysis in downstream applications. As Lysis-B Reagent is based on a Tris buffer system, Tris-HCl buffers are recommended for subsequent protein purification.
When extractions or single-step isolations are necessary in the acid pH range, simply include pepstatin along with cOmplete Mini EDTA-free tablets to ensure aspartic (acid) protease inhibition.
It has been used for cell incubation in the transient transfection assay, detection of expressed protein and western blot analysis.
Note: cOmplete Mini EDTA-free tablets (included in the kit) are employed to stabilize those protein extracts where activity of metal-containing proteins must not be affected. Since EDTA interferes with IMAC (immobilized metal 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
Time required: 10 minutes (bacterial lysis at room temperature).
Number of extractions: 20 (5ml each).
Effective against: Serine proteases, cysteine proteases (but not metalloproteases).
Note: Each cOmplete, Mini, EDTA-free tablet is concentrated enough to inhibit proteases in 5ml of extract.
Number of extractions: 20 (5ml each).
Effective against: Serine proteases, cysteine proteases (but not metalloproteases).
Note: Each cOmplete, Mini, EDTA-free tablet is concentrated enough to inhibit proteases in 5ml of extract.
- Saves time by completely and gently lysing bacteria in just 10 minutes, without sonication.
- Produces significantly higher yields of extracted proteins 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).
Packaging
1 kit containing 2 components
1 kit containing 2 components. Each tablet is sufficient for 5 mL of extract.
Preparation Note
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
cOmplete is a trademark of Roche
Kit Components Only
Product No.
Description
- Lysis-B reagent for protein extraction from bacterial cells, 100 ml 2x concentrated
- 20 cOmplete Mini EDTA-free Protease Inhibitor Cocktail Tablets in EASYpacks (foil blisters)
Regulatory Information
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