Recombinant Mouse Ubiquitin-Like Protein Isg15 (ISG15) Protein (His)

Name: Recombinant Mouse Ubiquitin-Like Protein Isg15 (ISG15) Protein (His)
Brand: Beta LifeScience
SKU: BLC-06493P
Availability: InStock

Greater than 85% as determined by SDS-PAGE.

Collections: High-quality recombinant proteins, Other recombinant proteins

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Product Overview

Description	Recombinant Mouse Ubiquitin-Like Protein Isg15 (ISG15) Protein (His) is produced by our Mammalian cell expression system. This is a full length protein.
Purity	Greater than 85% as determined by SDS-PAGE.
Uniprotkb	Q64339
Target Symbol	ISG15
Species	Mus musculus (Mouse)
Expression System	Mammalian cell
Tag	C-10His
Target Protein Sequence	MAWDLKVKMLGGNDFLVSVTNSMTVSELKKQIAQKIGVPAFQQRLAHQTAVLQDGLTLSSLGLGPSSTVMLVVQNCSEPLSILVRNERGHSNIYEVFLTQTVDTLKKKVSQREQVHEDQFWLSFEGRPMEDKELLGEYGLKPQCTVIKHLRLRGG
Expression Range	1-155aa
Protein Length	Full Length
Mol. Weight	19.2 kDa
Research Area	Cell Biology
Form	Liquid or Lyophilized powder
Buffer	Liquid form: default storage buffer is Tris/PBS-based buffer, 5%-50% glycerol. Lyophilized powder form: the buffer before lyophilization is Tris/PBS-based buffer, 6% Trehalose, pH 8.0.
Reconstitution	Briefly centrifuged the vial prior to opening to bring the contents to the bottom. Reconstitute protein in deionized sterile water to a concentration of 0.1-1.0 mg/mL. It is recommended to add 5-50% of glycerol (final concentration) and aliquot for long-term storage at -20°C/-80°C. The default final concentration of glycerol is 50%.
Storage	1. Store at -20°C/-80°C upon receipt, aliquoting is necessary for mutiple use. 2. Avoid repeated freeze-thaw cycles. 3. Store working aliquots at 4°C for up to one week. 4. In general, protein in liquid form is stable for up to 6 months at -20°C/-80°C. Protein in lyophilized powder form is stable for up to 12 months at -20°C/-80°C.
Notes	Repeated freezing and thawing is not recommended. Store working aliquots at 4°C for up to one week.

Target Details

Target Function	Ubiquitin-like protein which plays a key role in the innate immune response to viral infection either via its conjugation to a target protein (ISGylation) or via its action as a free or unconjugated protein. ISGylation involves a cascade of enzymatic reactions involving E1, E2, and E3 enzymes which catalyze the conjugation of ISG15 to a lysine residue in the target protein. Its target proteins include SERPINA3G/SPI2A, JAK1, MAPK3/ERK1, PLCG1, TRIM25, STAT5A, MAPK1/ERK2 and globin. Isgylation of the viral sensor IFIH1/MDA5 promotes IFIH1/MDA5 oligomerization and triggers activation of innate immunity against a range of viruses, including coronaviruses, flaviviruses and picornaviruses. Can also isgylate: DDX58/RIG-I which inhibits its function in antiviral signaling response, IRF3 which inhibits its ubiquitination and degradation as well as EIF4E2 which enhances its cap structure-binding activity and translation-inhibition activity. Exhibits antiviral activity towards both DNA and RNA viruses, including influenza A and B virus, sindbis virus (SV) and herpes simplex type-1 (HHV-1). Plays a significant role in the control of neonatal Chikungunya virus (CHIKV) infection by acting as a putative immunomodulator of proinflammatory cytokines. Protects mice against the consequences of Chikungunya virus infection by downregulating the pathogenic cytokine response, often denoted as the cytokine storm. Plays a role in erythroid differentiation. The secreted form of ISG15 can: induce natural killer cell proliferation, act as a chemotactic factor for neutrophils and act as a IFN-gamma-inducing cytokine playing an essential role in antimycobacterial immunity. The secreted form acts through the integrin ITGAL/ITGB2 receptor to initiate activation of SRC family tyrosine kinases including LYN, HCK and FGR which leads to secretion of IFNG and IL10; the interaction is mediated by ITGAL.
Subcellular Location	Cytoplasm. Secreted.
Database References	KEGG: mmu:100038882 STRING: 10090.ENSMUSP00000082548 UniGene: PMID: 27882925 Murine Isg15 does not control Usp18 accumulation and IFN-alpha/beta signaling. PMID: 27193971 Extracellular ISG15 signals cytokine secretion through the LFA-1 integrin receptor (CD11a/CD18) in natural killer cells. PMID: 29100055 results demonstrate that ISG15 governs the dynamic functionality of mitochondria, specifically, OXPHOS and mitophagy, broadening its physiological role as an antiviral agent. PMID: 29077752 Loss of ISG15 expression and type I IFN signaling have transient effects on bacterial burden during M. tuberculosis infection in mice PMID: 28087453 ISG15 acts as an immunomodulator in the cornea and plays a critical role in controlling fungal keratitis. PMID: 28599020 USP18's ISG15 specificity is mediated by a small interaction interface. PMID: 28165509 Posttranslational protein modification by ISG15 plays an important role in antiviral defense PMID: 25805508 These studies provide evidence, ISG15 is elevated following TBI in mice, preceding MLCK elevation, development of BBB disruption, and cerebral edema. PMID: 25669448 Kruppel-like factor 9 (KLF9) is a haploinsufficient tumor suppressor in the ApcMin/+ mouse colon by suppressing expression of ISG15, an apoptosis-inhibiting cytokine. PMID: 26210742 Propose that Isg15-dependent degradation of p53 represents an alternative mechanism of controlling p53 protein levels, and, thus, it is an attractive pathway for drug discovery. PMID: 24844324 Embryo mortality occurs in pregnant Isg15(-/-) mice, is exacerbated by environmental insults like maternal hypoxia, and might result from impaired early decidualization, vascular development, and formation of the labyrinth. PMID: 25505199 Enhanced resistance upon influenza B infection in USP18(C61A/C61A) mice was completely reversed in USP18(C61A/C61A) mice, which additionally lack ISG15, providing evidence that the observed reduction in viral titers is ISG15 dependent PMID: 25605921 Authors found that ISG15 protects mice from influenza A virus and Sendai virus induced lethality by a conjugation-dependent mechanism in both of these models. PMID: 25320315 study demonstrated that ISG15 had an anti-apoptotic effect on the beta cell line MIN6; conclusion is consistent with previous reports that ISG15 has a protective role PMID: 25031023 The ISG15 conjugation system represents a critical innate response mechanism in cardiomyocytes to fight the battle against invading pathogens, limiting inflammatory cardiomyopathy, heart failure, and death. PMID: 25165091 ISG15 does not appear to play a key role in IFN-beta-mediated C2C12 myoblast cell fusion PMID: 23750257 Interferon-stimulated gene 15 (ISG15) mitigates MNV-1 replication. PMID: 24899198 Protein interferon-stimulated gene 15 conjugation delays but does not overcome coronavirus proliferation in a model of fulminant hepatitis. PMID: 24648452 these results indicate an essential role of ISG15 in the cellular immune antiviral response PMID: 24137104 Isg15 regulates the expression of proteins that directly affect the HCV replication. The pro-viral effect of Isg15 in HCV replication might be mediated through these effector proteins. PMID: 24416772 Activation of double-stranded RNA-activated protein kinase (PKR) by interferon-stimulated gene 15 (ISG15) modification down-regulates protein translation PMID: 23229543 vaccination against ISG15 results in significant CD8-mediated reductions in both primary and metastatic mammary tumor burden. PMID: 22057675 Data show that the HerC6 N-terminal RCC1-like domain (RLD) allows ISG15 conjugation. PMID: 22272257 mHERC6 is essential for endogenous murine ISGylation and thus represents the dominant ISG15 E3 ligase in mice. PMID: 22138649 These findings suggest that ISG15 plays an anti-dengue virus and West Nile virus function via protein ISGylation. PMID: 21992229 ISG15 is induced during late erythroid differentiation. PMID: 22022510 increased protein ISGylation might be an endogenous neuroprotective adaptation to minimize poststroke brain damage PMID: 21847135 expression of the ISG15 is elevated in A-T cells derived from various A-T patients, as well as in brain tissues derived from the ATM knockout mice and A-T patients, suggesting that ATM negatively regulates the ISG15 pathway PMID: 21298066 In addition to cellular ISGylation targets, recent reports implicate viral proteins as targets of ISG15 modification; these studies provide fresh insights into the antiviral mechanisms of ISG15.[Review] PMID: 20946978 replication of hepatitis C virus was inhibited by overexpression of ISG15 PMID: 20810994 Data show that contrary to earlier reports, up to 50% of the fetuses die between 7.5 and 12.5 d post coitum (dpc) in Isg15(-/-) mothers when mated to Isg15(-/-) fathers. PMID: 20660068 Ectopic expression of the Herc5-mediated ISGylation system could distinctly potentiate IFN-beta-induced antiviral effects against influenza A virus. PMID: 20385878 The authors demonstrate that influenza B virus NS1 protein potently antagonizes human but not mouse ISGylation, a property dependent on B/NS1 binding the N-terminal domain of human but not mouse ISG15. PMID: 20219937 UBP43 (USP18) specifically removes ISG15 from conjugated proteins PMID: 11788588 lipopolysaccharide activation of expression via interferon regulatory factor 3 PMID: 11854279 UBP43 plays a critical role in maintaining the homeostatic balance of ISG15-conjugated protein, and that regulation of cellular levels of ISG15 protein modification is essential for brain cell function. PMID: 12208842 Isg15 is one of few gene products identified in murine implantation sites to require presence of the conceptus and not simply differentiation of the stroma. PMID: 12810567 results suggest that the ubiquitin family protein ISG15/UCRP has novel functions in neutrophil-mediated immune mechanisms PMID: 12963022 Ubc8 is a major ISG15-conjugating enzyme responsible for protein ISGylation upon interferon stimulation PMID: 15485925 ISG15 is dispensable for STAT1 and interferon signaling PMID: 16024773 These data indicate that ISGylation targets proteins found in several fundamentally important cellular pathways and will contribute to understanding the physiologic role of interferon-induced ISG15 and ISG15 conjugation. PMID: 16139798 ISG15 expression protected against Sindbis virus-induced lethality and decreased Sindbis virus replication in multiple organs without inhibiting the spread of virus throughout the host. PMID: 16254333 The phenotype of UBP43(-/-) mice was not rescued by the absence of ISG15, as evident from unchanged mortality, neurological symptoms, and occurrence of hydrocephalus. PMID: 16314524 Results show that Ubp43 negatively regulates type I interferon signaling independent of its isopeptidase activity towards ISG15. PMID: 16710296 Virus-mediated subversion of the antiviral response by proteolysis of IRF-3 is counteracted by induction of ISG15 expression. PMID: 16914094 ISG15 is a novel antiviral molecule with activity against both RNA and DNA viruses provides a target for the development of therapies against important human pathogens PMID: 17227866 Isg15 and its conjugates are present in implantation sites during mid to late gestation in mice. PMID: 17381847 Leishmania braziliensis infection triggered transcription and phosphorylation of STAT molecules and IFN-stimulated gene 15 PMID: 18490754 nitrosylation of ISG15 enhances target protein ISGylation PMID: 18606809

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Proteins are sensitive to heat, and freeze-drying can preserve the activity of the majority of proteins. It improves protein stability, extends storage time, and reduces shipping costs. However, freeze-drying can also lead to the loss of the active portion of the protein and cause aggregation and denaturation issues. Nonetheless, these adverse effects can be minimized by incorporating protective agents such as stabilizers, additives, and excipients, and by carefully controlling various lyophilization conditions.

Commonly used protectant include saccharides, polyols, polymers, surfactants, some proteins and amino acids etc. We usually add 8% (mass ratio by volume) of trehalose and mannitol as lyoprotectant. Trehalose can significantly prevent the alter of the protein secondary structure, the extension and aggregation of proteins during freeze-drying process; mannitol is also a universal applied protectant and fillers, which can reduce the aggregation of certain proteins after lyophilization.

Our protein products do not contain carrier protein or other additives (such as bovine serum albumin (BSA), human serum albumin (HSA) and sucrose, etc., and when lyophilized with the solution with the lowest salt content, they often cannot form A white grid structure, but a small amount of protein is deposited in the tube during the freeze-drying process, forming a thin or invisible transparent protein layer.

Reminder: Before opening the tube cap, we recommend that you quickly centrifuge for 20-30 seconds in a small centrifuge, so that the protein attached to the tube cap or the tube wall can be aggregated at the bottom of the tube. Our quality control procedures ensure that each tube contains the correct amount of protein, and although sometimes you can't see the protein powder, the amount of protein in the tube is still very precise.

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