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71092 Millipore

PopCulture Reagent

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Properties

Related Categories Molecular Biology, Native Protein Sample Preparation, Protein Extraction, Proteomics
Quality Level   100
mfr. no.   Novagen®
storage condition   OK to freeze
storage conditions   +15C to +30C
shipped in   ambient

Description

General description

PopCulture 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

Packaging

15, 250, 75 ml in Plastic ampoule

Warning

Toxicity: Irritant (B)

Other Notes

1. Grabski, A., Drott, D., Handley, M., Mehler, M. and Novy, R. (2001). inNovations13, 1–4.

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

Legal Information

This product is covered by EP Patent 1,432,822 owned by EMD Chemicals Inc. or its Affiliates.

Benzonase is a registered trademark of Merck KGaA, Darmstadt, Germany

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