His-Tag, Recombinant, Western Blot Positive Control (6-Histidine)

Western Blot Positive Control for H5099-15C.\n\nRecombinant DNA technology allows the addition of short pieces of well-defined tags, "peptides" or proteins at the amino or c-terminus of target genes, which can provide 'affinity handles' designed to bind specific matrices. Therefore, tags enables a selective identification and purification of the protein of interest. Poly-His (His x 6) containing proteins can be purified by immobilized metal ion affinity chromatography (IMAC), making use of its high affinity for transition metal ions (1-4). This purification system eliminates the harsh conditions required to elute protein from ligand affinity columns. Numerous recombinant fusion proteins have been engineered with histidine tags to allow detection, isolation and purification of fusion proteins (1-4). Anti-poly may be used to identify the expression of a poly-histidine fusion protein in bacteria, bacterial lysates or cells and tissues transfected with a poly-histidine fusion protein expression vectors.\n\nSource:\nHis-tag-fusion protein., expressed as N-terminal fusion protein of ~10kD in E. coli.\n\nMolecular Weight:\n~10kD\n\nApplications:\nSuitable for use in Western Blot. Not suitable for use in ELISA. Other applications not tested.\n\nRecommended Dilutions:\nWestern Blot: 10ul/lane. The protein migrates as ~9-10kD band. It will be necessary to have proper size gel (e.g., 10% or 4-10% for proper resolution of small protein. SDS may crystallize in cold conditions. It should redissolve by warming and light mixing at room temperature. It should be heated once prior to loading on gels. If the product has been stored for several weeks, then it may be preferable to add 5ul of fresh 2x sample buffer per 10ul of the H5099-15D solution prior to heating and loading on gels. This preparation is not biologically active.\nOptimal dilutions to be determined by the researcher.\n\nStorage and Stability:\nAliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for at least 6 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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