C8740 does not interfere with downstream applications such as 6X HisTag affinity tag purification, or detection on a Western Blot with an anti-6X His antibody.
If the intention is to conjugate a protein to a 6X HisTag, the protein should first be purified from the lysate before conjugation. C8740 will not interfere with a conjugation protocol after the protein of interest has been isolated from the lysate.
C8740
CelLytic™ B Cell Lysis Reagent
For bacterial cell lysis, 10× concentrate
Synonym(s):
bacterial cell lysis reagent
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About This Item
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Quality Level
form
liquid
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Application
TM[="CelLytic"] B Cell Lysis Reagent has been used for lysing E.coli 5-helix bundle protein.[1] It is also suitable for lysing hepatitis B X-interacting protein (HBXIP)[2] and E. coli TG1 cells.[3]
A proprietary, non-denaturing formulation of zwitterionic detergents used for the lysis of bacterial cells and extraction of recombinant proteins.
Features and Benefits
- Higher protein extraction efficiency than traditional methods such as sonication and lysozyme
- Scalable for 1 to 25 grams of bacterial cell paste
- No interference with downstream applications such as affinity chromatography, IP, and Western blotting
- Compatible with protease inhibitors, inhibitor cocktails, chaotropes, salts, chelating agents and reducing agents
Other Notes
10X concentrate of detergent blend contains no buffering components, giving the researcher flexibility to integrate the lysis reagent into their desired system. Also amenable to automation protocols for recombinant protein extraction and purification.
Legal Information
Covered by US Patent No 7,282,475 B2 and are sold for research use only. Commercial use requires addtional licenses.
CelLytic is a trademark of Sigma-Aldrich Co. LLC
Signal Word
Warning
Hazard Statements
Precautionary Statements
Hazard Classifications
Eye Irrit. 2 - Skin Irrit. 2
Storage Class Code
12 - Non Combustible Liquids
WGK
WGK 3
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Personal Protective Equipment
dust mask type N95 (US), Eyeshields, Gloves
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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Can this product safe to be used for downstream crosslinking stains such as a 6x HIsTag?
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When using the CelLytic™ B lysis reagent, do I need to make a dilution prior to measuring protein by Bradford assay?
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When using the CelLytic™ B lysis reagent (B7435, B7310, C8740, and contained in kits CB0050 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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What is the Department of Transportation shipping information for this product?
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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.
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What bacterial cells can be used with CelLytic B?
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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.
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Can I use the CelLytic™ B cell lysate in an activity assay?
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CelLytic B contains a zwitterionic detergent so the protein is active and can be used in protein activity assays.
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What is the difference between CelLytic B (Product Nos. B7435, B7310, C8740) and CelLytic B Plus (Product Nos. CB0050 and CB0500)?
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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.
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What is the difference between CelLytic B (Product No. B7435), CelLytic B 2x (Product No. B7310) and CelLytic B 10x (Product No. C8740)?
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CelLytic B 2X has double the strength of CelLytic B for small volume extractions. This allows CelLytic B 2X to be used when a higher protein concentration or lower volume is required. The original formula, CelLytic B, requires 10-20 ml of the reagent for 1 gram of wet cell paste. Only 5 ml of the CelLytic B 2X reagent is required to lyse and extract protein from 1 gram of wet cell paste. CelLytic B 10X is provided as a concentrated mixture of the proprietary detergents found in CelLytic B, without any buffering component. This allows the user to customize the extraction reagent by choosing the detergent concentration and buffering components ideal for the extraction and purification of the protein(s) of interest. CelLytic B 10X is also suitable for the lysis of yeast and mammalian cells.
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Can I obtain protein from inclusion bodies with CelLytic B?
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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.
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Can I concentrate my sample that was made using CelLytic B?
1 answer-
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.
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Can I use CelLytic B with the maltose binding protein system (MBP System)?
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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.
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