71092
PopCulture® Reagent
Synonym(s):
E. coli culture medium protein extraction reagent
Select a Size
About This Item
form
liquid
Quality Level
manufacturer/tradename
Novagen®
storage condition
OK to freeze
shipped in
ambient
storage temp.
10-30°C
Related Categories
General description
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
Other Notes
2. Inouye, M., Arnheim, N. and Sternglanz, T. (1973). J. Biol. Chem.248, 7247.
Legal Information
Disclaimer
Storage Class
10 - Combustible liquids
wgk_germany
WGK 3
flash_point_f
Not applicable
flash_point_c
Not applicable
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
Already Own This Product?
Find documentation for the products that you have recently purchased in the Document Library.
Related Content
Traditionally, protein purification from E. coli consists of four distinct phases: harvest, bacterial cell lysis, lysate clarification and protein purification. Bacterial lysis typically requires several time-consuming, hands-on steps, such as freeze/thaw cycles and sonication. These harsh lysis techniques may negatively impact protein quality and contribute to sample-to-sample variability. To maintain protein activity and integrity, detergent-based lysis buffers are routinely used to avoid mechanical protein extraction methods. Regardless of the lysis method used, centrifugation is traditionally required to pellet unwanted cell debris and permit recovery of the clarified lysate. The final step, purification, is frequently performed using affinity media specific for expressed epitope tags. Agarose-based media have typically been used, either as a slurry in microcentrifuge tubes or packed into gravity-driven or spin columns. While easier to manipulate, columns are greatly affected by lysate consistency and carryover of cell debris, which can lead to clogging of the column frits.
E. coli transformation with poly-histidine tagged constructs represents a common vehicle for protein production. Initial screening of small-scale cultures is routinely performed to identify clones producing the highest amounts of protein with the desired form and function. As the demand for greater throughput at this level has increased, so has the need for process simplification and greater reproducibility. To meet this need, we have developed a condensed workflow that can be performed in a single device.
Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.
Contact Technical Service