For years, the use of polyhistidine tags (His-tags) has been a staple in the isolation of recombinant proteins in immobilized metal affinity chromatography experiments. Its use has been highly beneficial in increasing the protein purity of crude cell lysates. For some recombinant proteins, one consequence of the addition of the His tag is that it can affect protein function and stability.
Functional proteins are essential in elucidating their biological, kinetic, structural, and thermodynamic properties. In this study, we determined the effect of N-terminal 6xHis-TF-tagged Recombinant on the thermal stability of selected proteins by differential scanning fluorimetry and identified that His tag removal can have both beneficial and detrimental effects on their stability.
Species reactivity: Label
Published species: Label
Host/Isotype: Rabbit / IgG
Class: Recombinant polyclonal
Immunogen: Synthetic construction corresponding to amino acids HHHHHH
Purification: Protein A
Storage buffer: PBS
Contains: 0.09% sodium azide
Store at 4°C for the short term. For long-term storage, store at -20°C, avoiding freeze/thaw cycles.
Product Specific Information
Recombinant rabbit polyclonal antibodies are unique offerings from Thermo Fisher Scientific. They are made up of a selection of multiple different recombinant monoclonal antibodies, providing the best of both worlds: the sensitivity of polyclonal antibodies with the specificity of monoclonal antibodies, all delivered with the consistency only found in a recombinant antibody.
While functionally the same as a polyclonal antibody, recognizing multiple epitope sites on the target and producing higher detection sensitivity for low-abundance targets, a recombinant rabbit polyclonal antibody has a known mix of heavy and light chains. The exact population can be produced in each batch, circumventing the biological variability typically associated with polyclonal antibody production.
The 6x His tag is a synthetic oligopeptide consisting of 6 consecutive histidine residues (HHHHHH). The His tag is commonly expressed as a tag at the 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 by genetic engineering. By choosing a particular combination of epitope and recombinant protein, epitope tagging makes it possible to detect proteins for which no antibodies are available.
His-epitope tagged proteins containing a stretch of histidine residues at the carboxyl or amino terminus are used for purification of recombinant proteins by metal chelate chromatography due to the metal-binding properties of the histidine groups on His-tagged proteins. For example, the affinity of the histidine tag motif for Ni2+ by chelation is strong and selective enough to allow purification of the protein to homogeneity by affinity chromatography on Ni2+-NTA adsorbent. Your tag-specific antibodies are used to facilitate the detection or co-immunoprecipitation of tagged proteins.