Chemistry:NE-tag
The NE-tag is a synthetic peptide tag (NE tag) designed as an epitope tag for detection, quantification and purification of recombinant protein. This patented peptide sequence is composed of eighteen hydrophilic amino acids. This short peptide does not adopt any significant homology to any existing proteins found in nature. This synthetic NE peptide adopts random coil conformation and showing strong immunogenicity (computational prediction). This is advantageous to offer stringent specificity to the NE-tagged proteins, which are readily to be detected, quantitated, and purified.[1]
Detection
The NE-tag can be specifically detected using a monoclonal anti-NE detection antibody - an affinity-purified mouse immunoglobulin, IgG1, which specifically binds to NE-tagged proteins. This peptide-to-antibody conjugation is validated in Western blotting, immunoprecipitation (IP), immunocytochemistry (IHC), and affinity purification of NE fusion proteins (i.e. affinity column).[2][3]
Sequences
- The DNA sequence of NE-tag is as follows: 5' ACC AAA gAA AAC CCg CgT AgC AAC Cag gAA gAA AgC TAT gAT gAT AAC gAA AgC 3'
- The corresponding amino acid sequence is: TKENPRSNQEESYDDNES (Thr-Lys-Glu-Asn-Pro-Arg-Ser-Asn-Gln-Glu-Glu-Ser-Tyr-Asp-Asp-Asn-Glu-Ser)
Patents
References
- ↑ Ho, Philip WL.; Tse, Zero HM.; Liu, HF.; Lu, S.; Ho, Jessica WM.; Kung, Michelle HW.; Ramsden, David B.; Ho, SL. (2013). "Assessment of cellular estrogenic activity based on estrogen receptor-mediated reduction of soluble-form catechol-O-methyltransferase (COMT) expression in an ELISA-based system.". PLOS ONE 8 (9): e74065. doi:10.1371/journal.pone.0074065. PMID 24040167. Bibcode: 2013PLoSO...874065H.
- ↑ Tse, HF.; Ho, JC.; Choi, SW.; Lee, YK.; Butler, AW.; Ng, KM.; Siu, CW.; Simpson, MA. et al. (2013). "Patient-specific induced-pluripotent stem cells-derived cardiomyocytes recapitulate the pathogenic phenotypes of dilated cardiomyopathy due to a novel DES mutation identified by whole exome sequencing.". Human Molecular Genetics 22 (7): 1395–1403. doi:10.1093/hmg/dds556. PMID 23300193.
- ↑ Xu, EG.; Ho, Philip WL.; Tse, Zero HM.; Ho, SL.; Leung, KM. (2016). "Revealing ecological risks of priority endocrine disrupting chemicals in four marine protected areas in Hong Kong through an integrative approach.". Environmental Pollution 215: 103–112. doi:10.1016/j.envpol.2016.04.090. PMID 27179329.
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