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PopCulture® Reagent and Kits
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Protein extraction from E. coli cultures directly in the growth medium
PopCulture® Reagent* is a detergent-based concentrate that can be added directly to cultures of E. coli to effectively extract proteins without the need for cell harvest. Recombinant proteins can be directly screened in the crude extract, or purified by adding an affinity matrix, washing the matrix-target protein complex to remove spent culture medium and cellular contaminants, and eluting the purified protein from the matrix. The entire culturing, extraction, and purification process can be performed in the original culture tube or multiwell plate. This "in-media" protein screening or purification procedure can be adapted to high-throughput roboticprocessing of samples for proteomics research and any application that would benefit from the increased speed and convenience it provides. Successful purification of intact fusion proteins from total culture extracts has been demonstrated using His•Bind® and GST•Bind™ Resins (Grabski 2001). The use of His•Mag™ or GST•Mag™ Agarose Beads enables the entire procedure to be carried out in a single tube without the need for columns or centrifugation. Addition of rLysozyme™ Solution or the use of pLysS hosts increases the efficiency of protein extraction with the procedure. Benzonase® Nuclease may also be added to reduce the viscosity of the extract.
PopCulture Reagent is supplied as a ready-to-use Tris-buffered liquid concentrate that is stable at room temperature. |
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