A Novel Monoclonal Antibody for detection of a new patented synthetic “NE” fusion protein tag
A novel fusion protein tag, named “NE” - was designed as a tool for detection and purification of recombinant protein. This sequence is composed of 18 hydrophilic amino acids which does not share significant homology to any existing native protein. This synthetic peptide has least ordered secondary structure with strong immunogenicity. These properties offer stringent specificity to NE-tagged proteins, which are readily to be detected, quantitated, and purified.
Monoclonal anti-NE detection antibody is an affinity purified mouse immunoglobulin, IgG1, which binds to NE fusion proteins. This antibody is useful in a wide spectrum of applications including Western blotting, immunocytochemistry, immunoprecipitation, and affinity purification of NE fusion proteins when bound to solid support.
Applications: Protein detection, tracing, purification and quantification
NE Peptide Tag Sequence "TKENPRSNQEESYDDNES"
- Not Found in Nature
- Stringent antibody specificity
- Hydrophilic Amino Acids
- Good water solubility
- Strong Immunogenicity
- Minimal denaturation or inactivation of tagged proteins
- Minimal secondary structure
- 100% Random Coil
- Good antibody accessibility
- Patented in US, Europe and China
- US8927225, EP2328909, CN102203119
C84H128 N26 O40
Chemical Formula
2142.07 g/mol
Molecular Weight
pH 3.84
Isoelectric Point
(Net charge at pH 7: -4)
1280 cm-1M-1
Extinction coefficient
Good Surface Accessibility of “NE” peptide tag
Prediction method: Bent Petersen, Thomas Nordahl Petersen, Pernille Andersen, Morten Nielsen and Claus Lundegaard. A generic method for assignment of reliability scores applied to solvent accessibility predictions. BMC Structural Biology (2009) 9:51.
| A | B | C | D | E | F | G | H | I |
|---|---|---|---|---|---|---|---|---|
| 1 | T | E | 0.856 | 118.755 | -0.407 | 0.003 | 0.003 | 0.994 |
| 2 | K | E | 0.63 | 129.612 | -0.716 | 0.113 | 0.043 | 0.844 |
| 3 | E | E | 0.655 | 114.516 | -1.3 | 0.113 | 0.043 | 0.844 |
| 4 | N | E | 0.51 | 74.664 | -1.05 | 0.115 | 0.016 | 0.868 |
| 5 | P | E | 0.334 | 47.395 | -1.909 | 0.181 | 0.016 | 0.803 |
| 6 | R | E | 0.497 | 113.699 | -0.684 | 0.257 | 0.016 | 0.727 |
| 7 | S | E | 0.493 | 57.768 | -1.248 | 0.339 | 0.016 | 0.645 |
| 8 | N | E | 0.485 | 70.975 | -1.334 | 0.339 | 0.016 | 0.645 |
| 9 | Q | E | 0.422 | 75.298 | -0.615 | 0.43 | 0.016 | 0.555 |
| 10 | E | E | 0.552 | 96.452 | -0.64 | 0.455 | 0.046 | 0.498 |
| 11 | E | E | 0.554 | 96.714 | -0.378 | 0.455 | 0.046 | 0.498 |
| 12 | S | E | 0.456 | 53.42 | -1.265 | 0.354 | 0.048 | 0.598 |
| 13 | Y | B | 0.239 | 51.053 | -0.247 | 0.257 | 0.016 | 0.727 |
| 14 | D | E | 0.53 | 76.301 | -0.723 | 0.257 | 0.016 | 0.727 |
| 15 | D | E | 0.529 | 76.272 | -1.558 | 0.257 | 0.016 | 0.727 |
| 16 | N | E | 0.588 | 86.112 | -2.711 | 0.181 | 0.016 | 0.803 |
| 17 | E | E | 0.642 | 112.14 | -1.431 | 0.115 | 0.016 | 0.868 |
| 18 | S | E | 0.902 | 105.714 | -0.27 | 0.003 | 0.003 | 0.994 |
Column A: Amino acid number
Column B: Amino acid
Column C: Class assignment – B for buried or E for Exposed – Threshold: 25% exposure, but not based on RSA
Column D: Relative Surface Accessibility – RSA
Column E: Absolute Surface Accessibility
Column F: Z-fit score for RSA prediction
Column G: Probability for Alpha-Helix
Column H: Probability for Beta-Strand
Column I: Probability for Coil
Mouse monoclonal NE© antibody, clone 5F6
For detection of a novel fusion protein tag
Production of mouse monoclonal anti-NE antibody
Validated Applications
Western blotting
(protein detection & quantification)
Immunocytochemistry
(Intracellular localization)
Immunoprecipitation
(protein-protein interaction)
Publications
- Ho PWL, Tse ZHM, Liu HF, Lu S, Ho JWM, Kung MHW, Ramsden DB and Ho SL. 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. 2013; 8(9):e74065.
- Tse HF, Ho JC, Choi SW, Lee YK, Butler AW, Ng KM, Siu CW, Simpson MA, Lai WH, Chan YC, Au KW, Zhang J, Lay KW, Esteban MA, Nicholls JM, Colman A, Sham PC. 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. Hum Mol Genet. 2013; 22(7):1395-403.
- Xu EG, Ho PWL, Tser Z, Ho SL, Leung KM. Revealing ecological risks of priority endocrine disrupting chemicals in four marine protected areas in Hong Kong through an integrative approach. Environ Pollut.2016; 215:103-112.
How to incorporate NE fusion protein tag into your target protein
Simply substitute your desired restriction site (R/S) and cDNA sequence of your gene-of-interest (G.O.I.)
… Your primers are ready to order and proceed.
N-terminal tag
Forward Primer Builder
5’-R/S —gCCACC—ATG—ACCAAAgAAAACCCgCgTAgCAACCAggAAgAAAgCTATgATgATAACgAAAgC—XXXXXXXXXX (G.O.I.)-3’
NE sequence
Approximate Melting point: 68.4℃ ; G.C control: 48%
C-terminal tag
Reverse Primer Builder (reverse complement)
5’-R/S —TTA—gCTTTCgTTATCATCATAgCTTTCTTCCTggTTgCTACgCgggTTTTCTTTggT—Linker—XXXXXXXXXX (G.O.I.)-3’
NE sequence
Approximate Melting point: 68.4℃ ; G.C control: 48%
"We invite you to bring this new protein tag and antibody into your research!"
Research Team
Dr. David B. RAMSDEN, PhD
Dr. Matthew Chi-Ting LEUNG, PhD
Dr. Colin Siu-chi LAM, PhD
Dr. Hui-Fang LIU, PhD
| Address | Division of Neurology, Department of Medicine, The University of Hong Kong, Li Ka Shing Faculty of Medicine Building, 21, Sassoon Road, Hong Kong. |
| Telephone | 852-2255 3315 |
Contact Person
Dr Philip Wing-Lok Ho (何永樂), PhD
| hwl2002@hku.hk | |
| HKU Scholar Hub | http://hub.hku.hk/rp/rp00259 |
