Bacterial PE LB™

Product catalog summary

Introduction
Bacterial PE LB™ is designed for extracting soluble proteins and inclusion bodies from bacterial cells, improving upon traditional lysozyme-based lysis. It facilitates the removal of nucleic acids during cell lysis, reducing viscosity and allowing effective clarification with lower centrifugal force. The kit includes protocols for spheroplast formation and protein extraction, utilizing organic buffering agents, mild non-ionic detergents, and a proprietary combination of salts and agents.

Applications
The product is suitable for preparing spheroplasts, lysing bacterial cells, extracting proteins, and isolating inclusion bodies. It is compatible with various downstream applications such as chromatography, gel electrophoresis, and protein folding procedures.

Compatibility
Bacterial PE LB™ is compatible with most downstream applications, including chromatography and electrophoresis, and can be used with the NI™ protein assay for protein estimation.

Items Supplied
The kit includes Bacterial PE LB™ in various volumes, Bacterial Suspension Buffer, and PE LB™ Lysozyme. Additional volumes of Bacterial PE LB™ Buffer can be purchased separately.

Storage Conditions
The kit should be stored at 4°C, with PE LB™ Lysozyme stored at -20°C. It remains stable for one year under recommended conditions.

Additional Items Needed
Users will need a centrifuge, test tubes, and an incubator. Additional Bacterial PE LB™ Buffer may be required for certain applications.

Preparation Before Use
Depending on the application, DTT and EDTA may be added to the Bacterial PE LB™ to a final concentration of 5mM. If divalent metal ions are necessary, EDTA should be omitted.

Protocols
A. Protein Extraction with Concurrent Removal of Nucleic Acids
1. Pellet bacterial cells and suspend in Bacterial PE LB™ Buffer.
2. Add PE LB™ Lysozyme and incubate.
3. Vortex and centrifuge to obtain clear lysate.

B. Protein Extraction with Spheroplast Formation
1. Pellet cells and suspend in Bacterial Suspension Buffer.
2. Add PE LB™ Lysozyme, incubate, and centrifuge to obtain spheroplasts.
3. Lyse spheroplasts and centrifuge to obtain lysate.

C. Isolation of Inclusion Bodies
1. After lysis, centrifuge lysate to collect inclusion bodies.
2. Wash inclusion bodies and prepare for solubilization and re-folding.

Related Products
Additional resources and related products can be found on the G-Biosciences website.

Catalog excerpts

G-Biosciences ♦ 1-800-628-7730 ♦ 1-314-991-6034 ♦ [email protected] A Geno Technology, Inc. (USA) brand name Bacterial PE LB™ Bacterial Protein Extraction Lysis Buffer (Cat. # 786-176, 786-177, 786-185, 786-186, 786-187, 786-188) think proteins! think G-Biosciences www.GBioscience

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INTRODUCTION ™ Bacterial PE LB has been developed for the extraction of soluble proteins and inclusion bodies from bacterial cells. It is a proprietary improvement on the lysozyme based lysis, which allows extraction of soluble proteins and concurrent removal of nucleic acids ™ (DNA & RNA) released during cell lysis. The Bacterial PE LB lysis eliminates viscosity build-up, allowing effective clarification with lower centrifugal force. This kit is provided with an optional protocol for the formation of spheroplast and removal lytic enzyme ™ (Lysozyme) prior to lysis and extraction of the bacterial...

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Bacterial PE LB Buffer Only Bacterial Suspension Buffer ™ PE LB Lysozyme (Cat# 786-042) STORAGE CONDITION The kit is shipped at ambient temperature. Upon arrival store the kit components at 4°C ™ except store PE LB Lysozyme at -20°C. Stable for 1 year when stored and used as recommended. ADDITIONAL ITEMS NEEDED • Centrifuge • Test tubes • Incubator ™ • Additional volume of the Bacterial PE LB Buffer may be purchased separately for downstream applications such as chromatography, dialysis, etc. PREPARATION BEFORE USE Depending on applications, DTT and EDTA may be added. Prepare an appropriate ™...

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PROTOCOLS A. Protein extraction with concurrent removal of nucleic acids 1. Pellet bacterial cells (bacterial culture, OD600 1.5-3.0) by centrifugation at 5,000x g for 10 minutes. ™ 2. Suspend the cell pellet in 5-10 volume of the Bacterial PE LB Buffer. For a 25µl cell ™ pellet (50-75mg wet weight), use 125-250µl Bacterial PE LB Buffer). 3. Vortex for 1 minute or until the cell suspension is homogeneous. Incubate the suspension for 5 minutes in cold. Vortex again to suspend the cells. ™ 4. Vortex the tube containing PE LB Lysozyme to mix the frozen suspension. Add an ™ ™ appropriate volume of...

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B. Protein extraction with spheroplast formation ™ Suitable when PE LB Lysozyme contamination is not acceptable. 1. Pellet bacterial cells (bacterial culture, OD600 1.5-3.0) by centrifugation at 5,000x g for 10 minutes. Suspend the cell pellet in 5-10 volume of the Bacterial Suspension Buffer (cell pellet size 25µl (50-75mg wet weight) use 125-250µl Bacterial Suspension Buffer). 2. Vortex for 1 minute or until the cell suspension is homogeneous. Incubate the o suspension for 5 minutes at 4 C. Vortex again to suspend the cells. ™ 3. Vortex the tube containing PE LB Lysozyme to mix the frozen suspension....

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RELATED PRODUCTS Download our Sample Preparation and Protease & Phosphatase Inhibitors, Enzyme & Assays Handbooks. http://info.gbiosciences.com/complete-protein-sample-preparation-handbook/ http://info.gbiosciences.com/protease-phosphatase-inhibitors-enzymes-assayhandbook For other related products, visit our website at www.GBiosciences.com or contact us. Last saved: 7/25/2016 CMH

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