The NE-tag is a synthetic

peptide Peptides are short chains of amino acids linked by peptide bonds. A polypeptide is a longer, continuous, unbranched peptide chain. Polypeptides that have a molecular mass of 10,000 Da or more are called proteins. Chains of fewer than twenty am ...

tag (NE tag) designed as an

epitope An epitope, also known as antigenic determinant, is the part of an antigen that is recognized by the immune system, specifically by antibodies, B cells, or T cells. The part of an antibody that binds to the epitope is called a paratope. Although e ...

tag for detection, quantification and purification of recombinant protein. This patented peptide sequence is composed of eighteen

hydrophilic A hydrophile is a molecule or other molecular entity that is attracted to water molecules and tends to be dissolved by water.Liddell, H.G. & Scott, R. (1940). ''A Greek-English Lexicon'' Oxford: Clarendon Press. In contrast, hydrophobes are n ...

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 Immunogenicity is the ability of a foreign substance, such as an antigen, to provoke an immune response in the body of a human or other animal. It may be wanted or unwanted: * Wanted immunogenicity typically relates to vaccines, where the injecti ...

(computational prediction). This is advantageous to offer stringent specificity to the NE-tagged proteins, which are readily to be detected, quantitated, and purified.

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 The western blot (sometimes called the protein immunoblot), or western blotting, is a widely used analytical technique in molecular biology and immunogenetics to detect specific proteins in a sample of tissue homogenate or extract. Besides detec ...

immunoprecipitation Immunoprecipitation (IP) is the technique of precipitating a protein antigen out of solution using an antibody that specifically binds to that particular protein. This process can be used to isolate and concentrate a particular protein from a sam ...

(IP),

immunocytochemistry Immunocytochemistry (ICC) is a common laboratory technique that is used to anatomically visualize the localization of a specific protein or antigen in cells by use of a specific primary antibody that binds to it. The primary antibody allows vis ...

(IHC), and

affinity purification Affinity chromatography is a method of separating a biomolecule from a mixture, based on a highly specific macromolecular binding interaction between the biomolecule and another substance. The specific type of binding interaction depends on the ...

of NE fusion proteins (i.e. affinity column).

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

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References

{{Protein tag Peptides