Bacterial PE LB™
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Bacterial PE LB™ - 1

G-Biosciences ♦ 1-800-628-7730 ♦ 1-314-991-6034 ♦ technical@GBiosciences.com 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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Bacterial PE LB™ - 3

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...

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Bacterial PE LB™ - 4

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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Bacterial PE LB™ - 5

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...

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Bacterial PE LB™ - 6

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...

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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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