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CB0050 Sigma-Aldrich

CelLytic B Plus Kit

For bacterial lysis

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Description

General description

The CelLytic B Plus Kit is designed to efficiently lyse cells and extract proteins from both Gram negative, and difficult to lyse, Gram positive bacteria. This is accomplished using the standard CelLytic B, a proprietary non-ionic detergent in concert with lysozyme, Benzonase®, and protease inhibitors. This complete kit takes the guesswork out of protein extraction from a variety of bacterial species.

Application

This kit is compatible with FLAG®, HIS-Select®, and glutathione S-transferase based affinity chroma­tography protein purification systems. The detergent included in the CelLytic B Lysis Reagent can be removed from the protein by dialysis or ammonium sulfate precipitation, if necessary. The final purity of the recombinant product is usually higher than that obtained from traditional extraction methods. This is because the crude protein solution containing the non-ionic detergent eliminates much of the non-specific binding that occurs during chromatography. The mild detergent does not interfere with many enzyme assays or protein assays.

Features and Benefits

• Lyse Gram positive and Gram negative bacteria
• More efficient than sonication
• Compatible with affinity purification
• Isolate inclusion bodies for subsequent solubilization
• Non-denaturing cell lysis preserves protein function

Other Notes

The CelLytic B Plus Kit contains all of the reagents and chemicals necessary to lyse both Gram-negative and Gram-positive bacteria. This kit also includes protease inhibitors to help prevent the proteolytic breakdown of proteins.

Legal Information

Benzonase® Nuclease is manufactured by Merck KGgA, Darmstadt, Germany and its affiliates. Nycomed Pharma A/S (Denmark) claims worldwide patent rights to Benzonase Nuclease, which are licensed exclusively to Merck KGaA, Darmstadt, Germany.

Covered by US Patent No 7,282,475 B2 and are sold for research use only. Commercial use requires addtional licenses.

Benzonase is a registered trademark of Merck KGaA, Darmstadt, Germany

CelLytic is a trademark of Sigma-Aldrich Co. LLC

FLAG is a registered trademark of Sigma-Aldrich Co. LLC

HIS-Select is a registered trademark of Sigma-Aldrich Co. LLC

Kit component also available separately

Description

Product #

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CelLytic B Cell Lysis Reagent, For bacterial cell lysis, standard strength SDS B7435
Benzonase® Nuclease, ≥250 units/μL, ≥90% (SDS-PAGE), recombinant, expressed in E. coli, buffered aqueous glycerol solution SDS E1014
Protease Inhibitor Cocktail, for use in purification of Histidine-tagged proteins, DMSO solution SDS P8849
Lysozyme from chicken egg white, 10 mg/mL SDS L3790
Safety & Documentation

Safety Information

Symbol 
GHS07  GHS07
Signal word 
Warning
Hazard statements 
Precautionary statements 
RIDADR 
NA 1993 / PGIII
Flash Point(F) 
Not applicable
Flash Point(C) 
Not applicable

Frequently Asked Questions

Which document(s) contains shelf-life or expiration date information for a given product?
If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis.
How do I get lot-specific information or a Certificate of Analysis?
The lot specific COA document can be found by entering the lot number above under the "Documents" section.
How do I find price and availability?
There are several ways to find pricing and availability for our products. Once you log onto our website, you will find the price and availability displayed on the product detail page. You can contact any of our Customer Sales and Service offices to receive a quote. USA customers: 1-800-325-3010 or view local office numbers.
What is the Department of Transportation shipping information for this product?
Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.
What bacterial cells can be used with CelLytic B?
CelLytic B Reagent by itself (no lysozyme or benzonase) was optimized for E. coli strain BL21.It also works with other common E. coli strains such as DH5?, JM109, and HB 101. The CelLytic B Reagent may be used on other similar bacterial cells. The CelLytic B Plus Kit is compatible with many more difficult to lyse Gram positive bacteria such as Bacillus and Streptococcus.
What is the difference between CelLytic B (Product Nos. B7435, B7310, C8740) and CelLytic B Plus(Product Nos. CB0050 and CB0500)?
CelLytic B Plus contains CelLytic B, Product No. B7435 as well as lysozyme, benzonase, and a protease inhibitor cocktail for a complete reagent for cell lysates for protein isolation.
What is the difference between CelLytic B Plus Kits, Products CB0050 and CB0500?
Product No. CB0050 and CB0500 provide reagents sufficient for processing 5 g or 50 g of cell paste, respectively.Fewer grams of cell paste can be processed if proteins are to be extracted from inclusion bodies.
Can I obtain protein from inclusion bodies with CelLytic B?
If the protein is found in inclusion bodies, you need to use an additional reagent, Product No. C5236, CelLytic IB to solubilize the proteins from the inclusion bodies.CelLytic B is used to isolate the inclusion bodies from the bacteria.CelLytic IB, Product No. C5236 can then be used to solubilize the proteins from the inclusion bodies.
Can I use the CelLytic B cell lysate in an activity assay?
CelLytic B contains a zwitterionic detergent so the protein is active and can be used in protein activity assays.
Can I concentrate my sample that was made using CelLytic B?
If concentrating a sample, be aware that concentration using filter concentrators may increase the detergent concentration significantly and cause issues in downstream applications.heoretically, ways to get around this are TCA precipitation, or diluting the extract below the CMC prior to concentration. This would be at least a 1/5 dilution, based on the following reference: Saito, S., and Tsuchiya, T., Characteristics of n-octyl beta-D-thioglucopyranoside, a new non-ionic detergent useful for membrane biochemistry. Biochem. J., 222(3), 829-832, 1984.
Can I use CelLytic B for eukaryotic cells?
CelLytic B was optimized for bacterial cells.It may lyse some mammalian cells. CelLytic B 10X is a better choice for the lysis of yeast and mammalian cells due to the increased detergent concentration.
Can I use CelLytic B with the maltose binding protein system (MBP System)?
CelLytic B cannot be used with the MBP system.We have used this in the maltose binding protein system, and know that it will NOT work in this system, since any and all detergents will destroy binding of protein to the resin.
When using the CelLytic B lysis reagent, do I need to make a dilution prior to measuring protein by Bradford assay?
When using the CelLytic B lysis reagent (B7435, B7310, C8740, and contained in kitsCB0050 and CB0500), it is best to use a 1:10 dilution in order to reduce the risk of interference with the Bradford Reagent. A 1:20 dilution would be even better, as long as the protein level remains within the standard curve.
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