Eye Irrit. 2 - Skin Irrit. 2
12 - Non Combustible Liquids
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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.
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.
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.
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.
CelLytic B contains a zwitterionic detergent so the protein is active and can be used in protein activity assays.
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.
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.
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 (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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