71092 PopCulture® Reagent

71092
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      Overview

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      71092-3
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          Plastic ampoule 15 ml
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          71092-4
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              Plastic ampoule 75 ml
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              71092-5
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                  Plastic ampoule 250 ml
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                  Description
                  OverviewPopCulture Reagent* is a buffered mixture of concentrated detergents formulated to extract proteins from E. coli cells directly in their culture medium. Using this method, cell culture, protein extraction and purification performed in the original culture tube or multiwell plate. PopCulture perforates the E. coli cell wall without denaturing soluble protein and protects protein from the pH extremes produced in high density culture media. Recombinant proteins can be assayed directly or purified by adding an affinity matrix, washing the matrix:target protein complex to remove culture medium and cellular contaminants and eluting the purified protein from the matrix. Purification of fusion proteins from total culture extracts has been demonstrated using both IMAC and GST affinity chromatography methods (1).

                  To further enhance the PopCulture purification procedure, lysozyme and/or Benzonase® Nuclease may be added. Lysozyme cleaves a peptidoglycan bond in the E. coli cell wall, enhancing cell lysis and increasing the yield of protein (1, 2). Proteins may be expressed in a host encoding T7 lysozyme (pLysS host) or exogenous lysozyme may be added after the PopCulture Reagent. Benzonase Nuclease may also be added to degrade endogenous nucleic acids that may interfere with purification due to high viscosity.

                  The PopCulture protein purification procedure is ideally suited to high throughput (HT) robotic processing of samples for proteomics research or any application that would benefit from the increased speed and convenience. The magnetic agarose capture resins (e.g. GST•Bind™ Magnetic Agarose Beads, His•Bind® Magnetic Agarose Beads) are optimal for HT applications since the entire procedure can be performed in a single tube without the need for columns or centrifugation.

                  *patent pending

                  Catalogue Number71092
                  Brand Family Novagen®
                  References
                  References1. Grabski, A., Drott, D., Handley, M., Mehler, M. and Novy, R. (2001). inNovations 13, 1–4.

                  2. Inouye, M., Arnheim, N. and Sternglanz, T. (1973). J. Biol. Chem. 248, 7247.
                  Product Information
                  DeclarationThis product is covered by EP Patent 1,432,822 owned by EMD Chemicals Inc. or its Affiliates.
                  Applications
                  Biological Information
                  Physicochemical Information
                  Dimensions
                  Materials Information
                  Toxicological Information
                  Safety Information according to GHS
                  Safety Information
                  Product Usage Statements
                  Storage and Shipping Information
                  Ship Code Shipped at Ambient Temperature or with Blue Ice
                  Toxicity Irritant
                  Storage +15°C to +30°C
                  Do not freeze Ok to freeze
                  Packaging Information
                  Transport Information
                  Supplemental Information
                  Specifications

                  Documentation

                  PopCulture® Reagent SDS

                  Title

                  Safety Data Sheet (SDS) 

                  PopCulture® Reagent Certificates of Analysis

                  TitleLot Number
                  71092

                  References

                  Reference overview
                  1. Grabski, A., Drott, D., Handley, M., Mehler, M. and Novy, R. (2001). inNovations 13, 1–4.

                  2. Inouye, M., Arnheim, N. and Sternglanz, T. (1973). J. Biol. Chem. 248, 7247.

                  Citations

                  Title
                • Andrew T. Anfora and Rodney A. Welch. (2006) DsdX is the second D-serine transporter in uropathogenic Escherichia coli clinical isolate CFT073. Journal of Bacteriology 188, 6622-6628.
                • Ilya G. Serebriiskii, et al. (2005) A combined yeast/bacterial two-hybrid system: development and evaluation. Molecular and Cellular Proteomics 4, 819-826.
                • Henry Nguyen, et al. (2004) An automated small-scale protein expression and purification screening provides beneficial information for protein production. Journal of Structural and Functional Genomics 5, 23-27.
                • User Protocols

                  Title
                  TB323 PopCulture® Reagent