Efficient, gentle lysis and extraction of E.coli and other bacterial cells.
B-PER Bacterial Protein Extraction Reagents are designed to extract soluble protein from bacterial cells without harsh chemicals or mechanical procedures like sonication.
These easy-to-use cell lysis reagents are nonionic detergent solutions that effectively disrupt cells and solubilize native or recombinant proteins without denaturation. Certain formulations include DNase I and Lysozyme enzyme supplements to improve yield of large molecular weight proteins that are difficult to purify. B-PER Reagents are compatible with the addition of standard protease inhibitors and with downstream protein purification methods, such as GST purification and His-tagged immobilized metal affinity chromatography (IMAC).
Highlights:
Ready to use – B-PER Reagents provide one-step E. coli cell lysis by a mild, nonionic detergent (proprietary) in your choice of Tris or phosphate buffer formulations
Fast and simple – just add B-PER Reagent to a bacterial pellet, shake for 10 minutes and recover soluble proteins after pelleting the cell debris
Excellent yields – recover both soluble and insoluble recombinant protein from bacterial lysates and purify inclusion bodies to near-homogeneous levels
Flexible – B-PER Reagents are suitable for any scale of protein extraction and are available in phosphate and 1X and 2X Tris formulations, with and without enzymes
Compatible – completely compatible with addition of protease inhibitor cocktails, and resulting protein extract can be used in protein assays, typical affinity purification methods (e.g., GST, 6xHis) and other applications
Convenient, Ready-to-use Formats:
Thermo Scientific B-PER Reagent Selection Guide.
B-PER Product
Composition and Suitable Applications
B-PER
Detergent in Tris buffer; no enzyme components
Bacterial lysis
Purification of affinity tagged proteins
B-PER II
(2X B-PER)
2X B-PER (detergent in Tris buffer; no enzyme components)
Bacterial lysis for low cell density
Purification of proteins having low expression levels
B-PER
(in Phosphate Buffer)
Detergent in sodium phosphate buffer; no enzyme components
Amine-free formulation for direct compatibility of lysate with amine-reactive labeling and crosslinking
B-PER with Enzymes
Tris buffered formulation with lysozyme and DNase enzymes
Improved cell membrane and DNA digestion for increased yields
Recovery of large molecular weight proteins
Recovery of insoluble proteins from inclusion bodies
B-PER Direct with Enzymes
Tris buffered formulation with lysozyme and DNase enzymes
Lysis of bacteria directly in cell culture media
Ideal for screening 96-well microplate samples
Product Details:
B-PER Bacterial Extraction Reagents are more effective than traditional sonication and typical homemade lysis buffers, many of which include detergents and components that interfere with downstream applications. B-PER Reagents are formulated in 20mM Tris buffer (pH 7.5) or phosphate buffer (Part No. 78266). They extract native and soluble recombinant proteins and yield lysates that are directly compatible with most downstream workflows such as electrophoresis, affinity purification, immunoprecipitation, protein interaction analysis, crosslinking and protein labeling. If necessary, the mild detergent components can be removed by dialysis or gel filtration (desalting columns).
Use Pierce Universal Nuclease for Cell Lysis with B-PER Products to reduce the viscosity of bacterial extracts and improve downstream applications by digesting and removing nucleic acids (DNA and RNA) from protein preparations. Using this nuclease helps to improve the separation of lysate pellet from supernatant, enhances filtration of the treated lysate, improves chromatography processing time and increases the overall protein yield.
Efficient and complete protein extraction with Thermo Scientific B-PER Reagent. E. Coli expressing green fluorescent protein (GFP) was extracted five times with B-PER Reagent or PBS/sonication. Equal amounts of each extract was by separated by SDS-PAGE and stained with coomassie dye. Extraction with B-PER Reagent was nearly complete after three rounds (Lanes 1-3). Analysis of GFP activity (below) confirms that nearly all GFP protein was extracted in the first three rounds with B-PER Reagent.
References:
Bonamy, G.M.C., et al. (2005) Mol. Endocrinol. 19(5): 1213-1230.
Chen, C., et al. (2004). J. Cell. Biol. 167(1): 161-170.
Dorsey, C., et al. (2003). Microbiology 149: 1227-1238.
Fan, R.S., et al. (2005) J. Biol. Chem. 280(25): 24212-24220.
Gringhuis, S., et al. (2005). Mol. Cell. Biol. 25(15): 6454-6463.
Hsiao, P., et al. (2003). Mol. Cell. Biol. 23(17): 6210-6220.
Hunger-Glaser, I., et al. (2003). J. Biol. Chem. 278(25): 22631-22643.
Kaufman, L., et al. (2004). J Am Soc Nephrol. 15: 1721-1730.
Keenan, R., et al. (2005). PNAS 102(25): 8887-8892.
Kirshner, J., et al. (2003). J. Biol. Chem. 278(50): 50338-50345.
Oltra, E., Pfeifer, I., Werner, R. (2003). Endocrinology 144(7): 3148-3158.
Pu, Y., et al. (2005). J. Biol. Chem. 280(29): 27329-27338.
Sakai, D., et al. (2006). Development 133: 1323-1333.
Tanabe, K., et al. (2005). Mol. Biol. Cell 16: 1617-1628.
