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Bacterial Cell Lysis

CelLytic B

This is a highly efficient (Figure 1 a and Figure 1 b ) yet gentle reagent for the extraction of proteins from bacteria (E. coli). This reagent is a proprietary formulation of zwitterionic detergents in 40 mM Trizma®-HCl (pH 8.0). Treatment of bacterial cells with CelLytic B results in rapid and efficient extraction of proteins that are suitable for affinity purification and analysis. CelLytic B is the method of choice for recombinant protein extraction and purification from E. coli. It was demonstrated to work with both the Anti-FLAG® and HIS-Select™ purification systems (Figure 2).

CelLytic B 2X has double the strength of CelLytic B for small volume extractions. Only 5 ml of the CelLytic B 2X reagent is required to lyse and extract protein from 1 gram of wet cell paste. 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.

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 his or her protein(s) of interest. CelLytic B 10X is also suitable for the lysis of yeast and mammalian cells.

Features & Benefits

  • Scalable based on the amount of bacterial cell paste
  • No interference with downstream applications such as affinity chromatography, immunoprecipitation, and Western blotting
  • With the addition of lysozyme, inclusion bodies are ready for resolubilization after two simple washes (using CelLytic IB, C5236)
  • Compatible with protease inhibitors, inhibitor cocktails, chaotropes, salts, chelating agents, and reducing agents

 


Product #  Product Name Package Size
B7435 CelLytic B Cell Lysis Reagent
(standard strength)
50 ml
500 ml
B7310 CelLytic B 2X Cell Lysis Reagent
(2x strength)
50 ml
250 ml
C8740 CelLytic B 10X Cell Lysis Reagent 10 ml
50 ml
100 ml

 Figure 1 a - One gram of E. coli cell paste was extracted using CelLytic B, lysozyme, or a combination of lysozyme and sonication. CelLytic B was used at a ratio of 10 ml per gram of cell paste. The lysozyme treatment was performed using 1 mg/ml lysozyme (L6876) and 10 mM EDTA (E5134) on ice for 15 minutes. Sonication time was 2 minutes on ice. Total protein extracted was determined by BCA assay.


 Figure 1 b - One gram of E. coli cell paste was extracted using CelLytic B, lysozyme, sonication, a combination of lysozyme and sonication, or a commercially available bacterial extraction reagent. CelLytic B, Competitor N, and Competitor P were all used at a ratio of 10 ml per gram of cell paste. The lysozyme treatment was performed using 1 mg/ml lysozyme (L6876) and 10 mM EDTA (E5134) on ice for 15 minutes. Sonication time was 2 minutes on ice. Total protein extracted was determined by BCA assay.


Figure 2 - SDS-PAGE gel of E. coli cell extracts prepared with CelLytic B used in affinity purification. Lanes 1 and 8: Low molecular weight markers (M3913). Lanes 2, 4 and 6: Crude cell extracts that contain GST, FLAG and HIS-tagged fusion proteins. Lanes 3, 5 and 7: GST, FLAG, and HIS-tagged fusion proteins after affinity purification.


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