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71092

Millipore

PopCulture® Reagent

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Synonym(s):
E. coli culture medium protein extraction reagent

form

liquid

Quality Level

manufacturer/tradename

Novagen®

storage condition

OK to freeze

shipped in

ambient

storage temp.

10-30°C

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This Item
C2978T942471187
Millipore

Millipore

71092

PopCulture® Reagent

CelLytic™ M Cell Lysis Reagent, Suitable for Mammalian cell lysis and protein solubilization.

Sigma-Aldrich

C2978

CelLytic M

TRI Reagent® For processing tissues, cells cultured in monolayer or cell pellets

Sigma-Aldrich

T9424

TRI Reagent®

manufacturer/tradename

Novagen®

manufacturer/tradename

-

manufacturer/tradename

-

manufacturer/tradename

Novagen®

storage condition

OK to freeze

storage condition

-

storage condition

-

storage condition

OK to freeze

shipped in

ambient

shipped in

-

shipped in

-

shipped in

ambient

storage temp.

10-30°C

storage temp.

room temp

storage temp.

2-8°C

storage temp.

10-30°C

Quality Level

100

Quality Level

200

Quality Level

200

Quality Level

100

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<TMSYMBOL></TMSYMBOL> Magnetic Agarose Beads, His•Bind<TMSYMBOL></TMSYMBOL> 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

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
NOVAGEN is a registered trademark of Merck KGaA, Darmstadt, Germany
POPCULTURE is a registered trademark of Merck KGaA, Darmstadt, Germany

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


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