Protein Refolding Kit, BioAssay(TM)

Produktübersicht

• Artikelname: Protein Refolding Kit, BioAssay(TM)
• Artikelnummer: USB-P9107-60
• Hersteller Artikelnummer: P9107-60
• Alternativnummer: USB-P9107-60-1
• Hersteller: US Biological
• Kategorie: Kits/Assays

Produktbeschreibung

Protein Refolding Kit, BioAssay(TM)is a screening kit that enables researchers to pinpoint the critical factors for refolding their protein in as little as 1 hour. Unlike many traditional methods, this kit employs a fractional factorial matrix design that allows the researcher to screen their specific protein in 15 different buffers quickly and easily. Researchers are able to examine a wider range of conditions all within a single experiment, simplifying the process of identifying the best buffer composition and method for the refolding of a given protein. The kit comes with enough buffer for 10 refolding experiments, as well as supplemental dithiothreitol and a Glutathione Redox System. Each buffer is available for individual purchase and is supplemented with the necessary Glutathione Redox System and/or DTT. Individual buffers come in 500ml and 1000ml amounts. Protein Refolding Kit, BioAssay(TM) is designed to help simplify the process of identifying the buffer composition and method best suited for protein refolding. For more than 20 years E. coli has proved to be a reliable host for the production of heterologous proteins. The well defined genetics, readily available host-vector systems, and established methods has made E. coli the first choice for the expression of recombinant proteins. Despite the history of successes, the expression of heterologous proteins the production of soluble functional protein remains unpredictable. Frequently, the over expression of a protein in E. coli results in the formation of insoluble inclusion bodies. The reasons for inclusion body formation are not fully known. Since translation is a slower process than protein folding, it is likely that the misfolding of translation intermediates plays some role. Posttranslational modification, such as glycosylation and lyposylation, are known to affect the secondary structure of proteins. In bacteria, these modifications are mostly absent. Further, the chemical environment in which translation occurs in the eukaryotic cell is different than that of the bacterial cell. Each of these factors contributes to varying degrees to how the nascent polypeptide folds, or in the case of recombinant protein expression, misfolds (1). Several approaches have been used to mitigate misfolding during the over expression of proteins in E. coli. These include: 1. Fusion of the target protein with a more soluble partner, typically a bacterial protein,, 2. Co-expression of folding catalysts and chaperones, 3. Expression under cultures conditions which reduce the translation rates or effect the intracellular environment, 4. Modification of the protein sequence. Each approach has advantages and disadvantages which must be weighed in light of the intended end-use of the target protein. Further, not all proteins respond favorably to any given approach. Again which approach is best suited to a given protein must be determined empirically and success in producing and recovering soluble active protein is not guaranteed. Both a bane and blessing, the formation of inclusion bodies renders the expressed protein unusable. The purification of a protein as an inclusion body is relatively simple, easily scalable for commercial applications, and in many cases, can stabilize the protein until a sufficient degree of purity is obtained. The challenge is that the protein must then be recovered from the insoluble particle. The recovery of soluble active protein from purified inclusion bodies requires the denaturation of the polypeptide and then its refolding to an active form. Many examples of proteins recovered from inclusion bodies are well known and used for both commercial and academic applications. There are well established methods for purifying inclusion bodies and solublizing the aggregated protein by denaturation. There is, however, no reliable method for predicting the conditions needed to refold the protein. Thus, the identification of the conditions needed to properly refold the protei