6x-His Tag Monoclonal Antibody (HIS.H8), eBioscience™, Invitrogen™

Description

• Description: This HIS.H8 monoclonal antibody reacts with the 6X polyhistidine (6X His) tag
• The H-H-H-H-H-H motif is frequently fused to the N- or C-terminus of proteins to facilitate purification (e.g., nickel affinity chromatography and immunoprecipitation) and/or detection
• This antibody recognizes the 6X His tag anywhere within the protein of interest (i.e., N-terminus, C-terminus, or internal)
• Applications Reported: This HIS.H8 antibody has been reported for use in immunoprecipitation and western blotting
• Applications Tested: This HIS.H8 antibody has been tested by immunoblotting of reduced cell lysate expressing a His-tagged protein at less than or equal to 1 μg/ mL
• It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest
• Purity: Greater than 90%, as determined by SDS-PAGE
• Aggregation: Less than 10%, as determined by HPLC
• Filtration: 0.2 μm post-manufacturing filtered
• The 6x His tag is a synthetic oligo peptide consisting of 6 consecutive histidine residues (HHHHHH)
• The His tag is commonly expressed as a tag at either N-or C-terminal regions of recombinant proteins to allow isolation or purification by immobilized metal affinity chromatography
• Epitope tagging is a technique in which a known epitope is fused to a recombinant protein using genetic engineering
• By choosing a particular epitope and recombinant protein combination, epitope tagging makes it possible to detect proteins for which no antibody is available
• His epitope tagged proteins that contain a stretch of histidine residues at the carboxyl or amino terminus are used for purification of recombinant proteins by means of metal chelate chromatography due to metal binding properties of histidine clusters on His tagged proteins
• For example, the affinity of the histidine-tag motif to Ni2+ by chelation is strong and selective enough to enable purification of the protein to homogeneity by affinity chromatography on a Ni2+-NTA adsorbent
• His tag-specific antibodies are used to facilitate detection or coimmunopreciptation of tagged proteins.