Difference between revisions of "Part:BBa K4673028"
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The his-tag is a tag commonly employed in the purification of recombinant proteins, comprising a sequence of six histidines(His-His-His-His-His-His). Importantly, with a molecular weight of less than 0.84kDa, his-tag is recognized for its minimal impact on the protein's biological and structural attributes, preserving the protein's native conformation. Moreover, it plays a crucial role in simplifying the protein purification procedures. According to the combination principle of the imidazole ring and divalent metal ions, the his-tag can be combined with the C-terminus or N-terminus of the target protein to form a special structure, thereby enhancing both the efficiency of target protein isolation and its subsequent detection. | The his-tag is a tag commonly employed in the purification of recombinant proteins, comprising a sequence of six histidines(His-His-His-His-His-His). Importantly, with a molecular weight of less than 0.84kDa, his-tag is recognized for its minimal impact on the protein's biological and structural attributes, preserving the protein's native conformation. Moreover, it plays a crucial role in simplifying the protein purification procedures. According to the combination principle of the imidazole ring and divalent metal ions, the his-tag can be combined with the C-terminus or N-terminus of the target protein to form a special structure, thereby enhancing both the efficiency of target protein isolation and its subsequent detection. | ||
| + | In our team’s project, 6X his-tag produced an affinity tag that functions as a specific gene incorporated into the pET-22b-LK vector. On account of its function of being detected by antibodies, this tagged peptide plays an important role in western-blotting and serves the purpose as an antigen for immune-recognition. Therefore, our team is able to identify the target protein from the bands with diverse molecular weights in the results of western-blotting while conducting both IPTG induction and oleate induction. | ||
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Latest revision as of 15:36, 10 October 2023
The his-tag can be detected by antibodies
The his-tag is a tag commonly employed in the purification of recombinant proteins, comprising a sequence of six histidines(His-His-His-His-His-His). Importantly, with a molecular weight of less than 0.84kDa, his-tag is recognized for its minimal impact on the protein's biological and structural attributes, preserving the protein's native conformation. Moreover, it plays a crucial role in simplifying the protein purification procedures. According to the combination principle of the imidazole ring and divalent metal ions, the his-tag can be combined with the C-terminus or N-terminus of the target protein to form a special structure, thereby enhancing both the efficiency of target protein isolation and its subsequent detection. In our team’s project, 6X his-tag produced an affinity tag that functions as a specific gene incorporated into the pET-22b-LK vector. On account of its function of being detected by antibodies, this tagged peptide plays an important role in western-blotting and serves the purpose as an antigen for immune-recognition. Therefore, our team is able to identify the target protein from the bands with diverse molecular weights in the results of western-blotting while conducting both IPTG induction and oleate induction.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]