The His-tagged expression system is widely used in the production of recombinant protein. The 6xHis tag on the expressed recombinant proteins allows for efficient coupling to Ni++ affinity resins and purification by single-step chromatography. The His-tag can be cleaved at sites recognized by enzymes such as thrombin and enterokinases to isolate the protein of interest.
Applications
Suitable for use in Immunoprecipitation, ELISA and Western Blot. Other applications have not been tested.
Recommended Dilution
Immunoprecipitation: 10ug/application ELISA: 0.1-1ug/ml Western Blot: 1-5ug/ml Optimal dilutions to be determined by the researcher.
Storage and Stability
May be stored at 4°C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and store at -20°C. Aliquots are stable for at least 12 months at -20°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.
Immunogen
Synthetic peptide containing six His residues (6xHis)
Form
Supplied as a liquid in PBS, pH 7.4, containing 0.09% sodium azide
Purity
Purified by Protein A affinity chromatography.
Specificity
Specific for 6xHis. On Western blots, it identifies a band at ~ 37kD. Reactivity has been confirmed with TS-His fusion protein, and is expected to react with all His-tagged proteins regardless of species.
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