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B-PER Protein Extraction Reagents


Efficient, gentle lysis and extraction of E.coli and other bacterial cells.

B-PER Bacterial Protein Extraction Reagents

Thermo Scientific B-PER Bacterial Protein Extraction Reagents are designed to extract soluble protein from bacterial cells without harsh chemicals or mechanical disruption.

These easy-to-use cell lysis reagents are nonionic detergent-based solutions that effectively disrupt cells and solubilize native or recombinant proteins without denaturation. B-PER Complete Bacterial Protein Extraction Reagent is an all-inclusive formulation that combines the lysis reagent with lysozyme and a universal nuclease to enable mild extraction from both gram-negative and gram-positive bacteria. Traditional Tris-buffered and phosphate-buffered formulations without enzymes are also offered. All B-PER Reagents are compatible with downstream applications, such as affinity chromatography (e.g., immobilized metal affinity chromatography, glutathione chromatography), SDS-PAGE, and protein quantification (e.g., Pierce BCA Protein Assay, Pierce 660nm Protein Assay). Depending on the particular application, protease inhibitors, salts, reducing agents, denaturants, and chelating agents may be added to the reagent.

Highlights:

  • Ready to use – one-step cell lysis of gram-positive and gram-negative bacteria using a mild, nonionic detergent (proprietary) in Tris or phosphate buffer formulations
  • Fast and simple – just add B-PER Reagent to a bacterial pellet, incubate with mixing for 10 to 15 minutes and recover soluble proteins after pelleting the cell debris
  • Convenient B-PER Complete Reagent contains lysozyme and a universal nuclease in a single formulation with 4°C storage
  • Excellent yields – recover recombinant proteins from bacterial lysates or 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 inhibitors, and the 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 Complete
  • Detergent in Tris buffer with Lysozyme and Universal Nuclease
  • Improved cell lysis and DNA digestion, thereby releasing soluble proteins and reducing viscosity to increase yields
  • Recovery of large molecular weight proteins (high MW proteins)
  • Cost-effective equivalent to Merck Millipore Bugbuster™ Master Mix
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 solubilization and DNA digestion
  • Will be discontinued Sept 2014; please use B-PER Complete
B-PER Direct with Enzymes
  • Tris buffered formulation with lysozyme and DNase enzymes
  • Lysis of bacteria directly in cell culture media
  • Will be discontinued Sept 2014; please use B-PER Complete

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 Tris or phosphate buffer at physiological pH. 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.

B-PER Complete provides ultimate convenience and performance, because it includes optimized concentrations of both lysozyme and universal nuclease and is stable for 1 year with 4°C storage. Lysozyme facilitates lysis by solubilizing bacterial cell walls; Pierce Universal Nuclease reduces the viscosity of bacterial extracts and improves downstream applications by digesting and removing nucleic acids (DNA and RNA). B-PER Complete is most efficient for frozen cells but has been validated and optimized to achieve high yields with both fresh and frozen cells from gram-positive and gram-negative bacteria. (For optimal performance with fresh gram-negative bacteria, supplementation with 1mM EDTA is required.) B-PER Complete is compatible with GST-fusion protein purification, unlike other formulations of lysis buffers which may inhibit enzyme function.

High-yield bacterial protein extaction with B-PER Complete

Protein yield comparison of two bacterial cell lysis reagents. E. coli ER2566/pLATE51-Klenow, ER2566/pGST-CC-StpA and ER2566/pGST-Syk cell pellets (0.5g), were resuspended in 2.5mL aliquots of B-PER Complete Reagent or EMD Millipore™ BugBuster™ Master Mix with gentle vortexing for 15 minutes at room temperature. Insoluble cell debris was removed by centrifugation at 16,000 x g for 20 min at 4°C. Protein yields (concentrations) for soluble fractions were determined using the Pierce BCA Protein Assay Kit (Part No. 23225).

  • C = control (E. coli lysate from before induction with IPTG)
  • T = total protein (initial lysate before removal of cell debris by centrifugation)
  • S = soluble protein (final clarified lysate of IPTG-induced culture)

 

B-PER Complete is compatible with fusion protein purification

Thermo Scientific B-PER Complete provides better compatibility with purification of 6xHis and GST fusion proteins. E. coli ER2566/pLATE51-Klenow, ER2566/pGST-CC-StpA and ER2566/pGST-Syk cell pellets cell pellets (0.5g), were resuspended in 2.5mL aliquots of B-PER Complete Reagent (B-PER) or EMD Millipore™ BugBuster™ Master Mix (Bug) with gentle vortexing for 15 minutes at room temperature. Insoluble cell debris was removed by centrifugation at 16,000 x g for 20 min at 4°C. 6xHis-Klenow protein was purified using Thermo Scientific HisPur Ni-NTA Agarose. GST-Syk and GST-StpA proteins were purified using Thermo Scientific Glutathione Agarose.

  • C = control (E. coli lysate from before induction with IPTG)
  • S = soluble protein (lysate after induction with 0.1mM IPTG)
  • E = elution fraction after protein purification

 

More Product Data

References:

  1. Bonamy, G.M.C., et al. (2005) Mol. Endocrinol. 19(5): 1213-1230.
  2. Chen, C., et al. (2004). J. Cell. Biol. 167(1): 161-170.
  3. Dorsey, C., et al. (2003). Microbiology 149: 1227-1238.
  4. Fan, R.S., et al. (2005) J. Biol. Chem. 280(25): 24212-24220.
  5. Gringhuis, S., et al. (2005). Mol. Cell. Biol. 25(15): 6454-6463.
  6. Hsiao, P., et al. (2003). Mol. Cell. Biol. 23(17): 6210-6220.
  7. Hunger-Glaser, I., et al. (2003). J. Biol. Chem. 278(25): 22631-22643.
  8. Kaufman, L., et al. (2004). J Am Soc Nephrol. 15: 1721-1730.
  9. Keenan, R., et al. (2005). PNAS 102(25): 8887-8892.
  10. Kirshner, J., et al. (2003). J. Biol. Chem. 278(50): 50338-50345.
  11. Oltra, E., Pfeifer, I., Werner, R. (2003). Endocrinology 144(7): 3148-3158.
  12. Pu, Y., et al. (2005). J. Biol. Chem. 280(29): 27329-27338.
  13. Sakai, D., et al. (2006). Development 133: 1323-1333.
  14. Tanabe, K., et al. (2005). Mol. Biol. Cell 16: 1617-1628.

Related Resources:

B-PER Reagent FAQ
Cell lysis and protein extraction technical guide

Related Products:

Other Cell Lysis Reagents
Protease and Phosphatase Inhibitors
Universal Nuclease for Cell LysisLysozymeDNase I
Fusion Protein Purification Resins and Kits


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
89821 B-PER Complete Bacterial Protein Extraction Reagent
Formulation: Proprietary detergent in 25mM Tris-HCl, pH 7.6, containing lysozyme and universal nuclease
Sufficient For: 50g cell paste
250mL Product Instructions for product #89821 B-PER Complete Bacterial Protein Extraction Reagent MSDS for product #89821 B-PER Complete Bacterial Protein Extraction Reagent Certificate of Analysis for product #89821 B-PER Complete Bacterial Protein Extraction Reagent $280.00

Add to cart
89822 B-PER Complete Bacterial Protein Extraction Reagent
Formulation: Proprietary detergent in 25mM Tris-HCl, pH 7.6, containing lysozyme and universal nuclease
Sufficient For: 100g cell paste
500mL Product Instructions for product #89822 B-PER Complete Bacterial Protein Extraction Reagent MSDS for product #89822 B-PER Complete Bacterial Protein Extraction Reagent Certificate of Analysis for product #89822 B-PER Complete Bacterial Protein Extraction Reagent $500.00

Add to cart
78243 B-PER Bacterial Protein Extraction Reagent
Formulation: Proprietary detergent in 20mM Tris buffer, pH 7.5
Sufficient For: 40g cell paste
165mL Product Instructions for product #78243 B-PER Bacterial Protein Extraction Reagent MSDS for product #78243 B-PER Bacterial Protein Extraction Reagent $156.00

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90084 B-PER Bacterial Protein Extraction Reagent
Formulation: Proprietary detergent in 20mM Tris buffer, pH 7.5
Sufficient For: 60g cell paste
250mL Product Instructions for product #90084 B-PER Bacterial Protein Extraction Reagent MSDS for product #90084 B-PER Bacterial Protein Extraction Reagent Certificate of Analysis for product #90084 B-PER Bacterial Protein Extraction Reagent $195.00

Add to cart
78248 B-PER Bacterial Protein Extraction Reagent
Formulation: Proprietary detergent in 20mM Tris buffer, pH 7.5
Sufficient For: 125g cell paste
500mL Product Instructions for product #78248 B-PER Bacterial Protein Extraction Reagent MSDS for product #78248 B-PER Bacterial Protein Extraction Reagent $282.00

Add to cart
78266 B-PER Reagent (in Phosphate Buffer)
Formulation: Proprietary detergent in 50mM sodium phosphate buffer, pH 7.5
Sufficient For: 125g cell paste
500mL Product Instructions for product #78266 B-PER Reagent (in Phosphate Buffer) MSDS for product #78266 B-PER Reagent (in Phosphate Buffer) $289.00

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78260 B-PER II Bacterial Protein Extraction Reagent (2X)
Formulation: 2X formulation of Part No. 78248.
Sufficient For: 125g cell paste
250mL Product Instructions for product #78260 B-PER II Bacterial Protein Extraction Reagent (2X) MSDS for product #78260 B-PER II Bacterial Protein Extraction Reagent (2X) $289.00

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90078 B-PER with Enzymes Bacterial Protein Extraction Kit
Formulation: Multi-component kit
Sufficient For: 60g cell paste

Kit Contents:
B-PER Reagent, 250mL
Lysozyme, 50mg/mL, 0.5mL
DNase I, 2500 U/mL, 0.5mL
250mL kit Product Instructions for product #90078 B-PER with Enzymes Bacterial Protein Extraction Kit MSDS for product #90078 B-PER with Enzymes Bacterial Protein Extraction Kit $282.00

Add to cart
90079 B-PER with Enzymes Bacterial Protein Extraction Kit
Formulation: Multi-component kit
Sufficient For: 125g cell paste

Kit Contents:
B-PER Reagent, 500mL
Lysozyme, 50mg/mL, 2 x 0.5mL
DNase I, 2500 U/mL, 2 x 0.5mL
500mL kit Product Instructions for product #90079 B-PER with Enzymes Bacterial Protein Extraction Kit MSDS for product #90079 B-PER with Enzymes Bacterial Protein Extraction Kit $502.00

Add to cart
90080 B-PER Direct Bacterial Protein Extraction Kit
Formulation: Multi-component kit
Sufficient For: 5 x 96-well plates of cultured cells

Kit Contents:
B-PER Direct Reagent, 50mL
Lysozyme, 50mg/mL, 0.5mL
DNase I, 2500 U/mL, 0.5mL
50mL kit Product Instructions for product #90080 B-PER Direct Bacterial Protein Extraction Kit MSDS for product #90080 B-PER Direct Bacterial Protein Extraction Kit $185.00

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Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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