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ISGylation-Specific Antibody Production Services

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Creative Biolabs provides customized ISGylation-specific antibody with high specificity and affinity based on phage display technology or hybridoma technology. Antibodies are produced by immunizing animals with a synthetic peptide corresponding to amino acids of the ISG15 protein. This antibody detects endogenous levels of both free and conjugated ISG15 protein and does not cross-react with other ubiquitin family members, including ubiquitin, SUMO-1, SUMO-2, SUMO-3, and NEDD8.

ISGylation is a post-translational modification that an ubiquitin-like protein, interferon-stimulated gene 15 (ISG15), also known as ubiquitin cross-reacting protein (UCRP), is covalently coupled to a lysine residue of cytoplasmic and nuclear target proteins. Expression of ISG15 is induced in response to type I interferons or lipopolysaccharide treatment and secreted from monocytes and lymphocytes with anti-viral activity. Similar to ubiquitin, ISGylation is a reversible enzymatic process, although the full mechanism has not been identified. ISGylation involves a stepwise enzymatic reaction which activated by E1 enzyme (UBE1L), conjugated by E2 enzyme (UbcH8), and ligated by E3 enzyme such as EFP/TRIM25 and HERC5. ISG15/UCRP is removed from proteins by the deconjugating enzyme UBP43/USP18. Unlike other ubiquitin-like molecules, ISG15 has not been identified in lower eukaryotes (yeasts, nematodes, insects, plants), indicating its role in specialized functions in higher eukaryotes.

In contrast to ubiquitination, ISGylation does not target proteins for degradation, rather ISGylation is induced by type I interferons. The target proteins are some of the type I IFN-induced proteins including MxA, Hup56, and PKR, and involve type I IFN signaling, such as PLCγ1, JAK1, ERK1/2, and STAT1. Several other targets are gradually found with the diverse cellular pathways, such as cell motility and intracellular protein trafficking, glycolysis, stress responses, and translation regulating. These findings implicate the functions of protein ISGylation in various biological pathways from type I IFN-induced immune responses to numerous fundamental cellular pathways. For instance, IFNα/β is induced by mycobacterial infection, and then stimulates the synthesis and secretion of ISG15. The secretion of ISG15 can thereby contribute to the production of IFNγ by activating NK cells. The immune system cells are then stimulated by the produced IFNγ. Dysregulation of the IFN signaling and the ISGylation system cause many diseases, including cancer and bacterial infection.

Monoclonal Mouse Antibody Production Fig. 1 Protein modification by ISG15. (Jeon Y J and Yoo H M 2010)

Monoclonal Mouse Antibody Production Fig. 2 ISGylation and its function in the antimycobacterial response. (Pohl C and Dikic I. 2012)

The antibodies against ISG15 protein produced by Creative Biolabs via High-Affi™ technology is applicable in various immunoassays, including ELISA, Western Blot, Immunohistochemistry (IHC), and immunoprecipitation. Meanwhile, excellent results of immunoassay have also proven the specificity and affinity of our ISGylation-specific antibodies.

Creative Biolabs is an expert in providing global customers with high-quality antibodies of diverse post-translational modification (e.g. phosphorylation, acetylation, glycosylation, methylation, ubiquitination, sumoylation, citrullination, prenylation, and fatty acylation). Based on years of research and development experience, our excellent scientists will use the most appropriate methods to provide the most satisfactory services for customers.

In addition to the ISGylation-specific antibody, Creative Biolabs also provides a comprehensive list of PTM-specific antibody production services of your choice.


Reference
Jeon Y J, Yoo H M, and Chung C H. (2010). “Isg15 and immune diseases”. Biochim Biophys Acta, 5: 485-496.
Pohl C and Dikic I. (2012). “Fighting mycobacteria through isgylation”. EMBO Rep, 13(10): 872-873.




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