Biology:HA-tag
The HA-tag is a protein tag derived from the human influenza hemagglutinin (HA) protein, which allows the virus to target and enter host cells. An HA-tag is composed of a peptide derived from the HA-molecule corresponding to amino acids 98-106, which can be recognized and selectively bound by commercially available antibodies. This makes HA a powerful tool in molecular biology, commonly included in expression vectors and in the production of recombinant proteins. [1] Like other epitope tags, HA-tag is small and generally does not alter the traits of proteins it is attached to.[2] As a result HA-tags are often used to identify protein-protein interactions or to detect protein expression, using Co-Immunoprecipitation or Western blot respectively.[3][4]
The HA-tag is not suitable for detection or purification of proteins from apoptotic cells since it is cleaved by Caspase-3 and / or Caspase-7 after its sequence DVPD, causing it to lose its immunoreactivity.[5] Labeling of endogenous proteins with HA-tag using CRISPR was recently accomplished in-vivo in differentiated neurons.[6]
Sequence
The DNA sequences for the HA-tag include: 5'-TAC-CCA-TAC-GAT-GTT-CCA-GAT-TAC-GCT-3' or 5'-TAT-CCA-TAT-GAT-GTT-CCA-GAT-TAT-GCT-3'. The resulting amino acid sequence is YPYDVPDYA (Tyr-Pro-Tyr-Asp-Val-Pro-Asp-Tyr-Ala).[7]
See also
References
- ↑ "Purification of a RAS-responsive adenylyl cyclase complex from Saccharomyces cerevisiae by use of an epitope addition method.". Mol Cell Biol 8 (5): 2159–65. 1988. doi:10.1128/mcb.8.5.2159-2165.1988. PMID 2455217.
- ↑ Kimple, Michelle E.; Brill, Allison L.; Pasker, Renee L. (2013-09-24). "Overview of Affinity Tags for Protein Purification". Current Protocols in Protein Science 73: Unit–9.9. doi:10.1002/0471140864.ps0909s73. ISSN 1934-3655. PMID 24510596.
- ↑ "Anti-HA Tag Antibody Updated 28/10/2021". Merck Millipore. https://www.merckmillipore.com/GB/en/product/Anti-HA-Tag-Antibody,MM_NF-05-904.
- ↑ "Overview of Epitope Tagging - US" (in en). https://www.thermofisher.com/us/en/home/life-science/antibodies/antibodies-learning-center/antibodies-resource-library/antibody-methods/epitope-tagging-overview.html.
- ↑ "The HA tag is cleaved and loses immunoreactivity during apoptosis". Nature Methods 4 (2): 107–8. February 2007. doi:10.1038/nmeth0207-107. PMID 17264856.
- ↑ "High Throughput, High Resolution Mapping of Protein Localization in Mammalian Brain by In Vivo Genome Editing". Cell 165 (7): 1803–1817. 2016. doi:10.1016/j.cell.2016.04.044. PMID 27180908.
- ↑ "Addgene: HA-tag" (in en). https://www.addgene.org/55182/.
Further reading
- "Purification of a RAS-responsive adenylyl cyclase complex from Saccharomyces cerevisiae by use of an epitope addition method". Molecular and Cellular Biology 8 (5): 2159–65. May 1988. doi:10.1128/MCB.8.5.2159. PMID 2455217.
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