ISG15 antibody

Catalog No. ABIN103335

Product Details anti-ISG15 Antibody

Target: ISG15 (ISG15 Ubiquitin-Like Modifier (ISG15))

Reactivity: Human

Host: Rabbit

Clonality: Polyclonal

Conjugate: This ISG15 antibody is un-conjugated

Application: Western Blotting (WB), ELISA, Immunohistochemistry (IHC)

Purpose: ISG15 Antibody

Cross-Reactivity (Details): Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Rabbit Serum.

Characteristics: Synonyms: rabbit anti-ISG15 Antibody, G1P2 antibody, IFI 15 antibody, IFI15 antibody, Interferon alpha inducible protein antibody, Interferon induced 15 kDa protein antibody

Purification: This product is an IgG fraction antibody purified from monospecific antiserum by a multi-step process which includes delipidation, salt fractionation and ion exchange chromatography followed by extensive dialysis against the buffer stated above.

Immunogen:

Immunogen: This purified antibody was prepared from rabbit serum after repeated immunizations with recombinant human ISG15 protein.

Immunogen Type: Recombinant Protein

Isotype: IgG

Application Details

Application Notes:

Immunohistochemistry Dilution: User Optimized

Application Note: This purified polyclonal antibody reacts with human ISG15 by western blot and ELISA. Although not tested, this antibody is likely functional in immunohistochemistry and immunoprecipitation.   This antibody using the specified conditions may recognize other prominent intrinsic bands (UBLs or conjugates), especially at lower dilutions.  An 18.5 kDa band corresponding to human ISG15 is detected.  IFNα or IFNß stimulated HeLa cell lysates can be used as a positive control.

Western Blot Dilution: 1:200 - 1:1,000

ELISA Dilution: 1:2,000 - 1:10,000

Other: User Optimized

Restrictions: For Research Use only

Handling

Format: Lyophilized

Reconstitution:

Reconstitution Volume: 100 μL

Reconstitution Buffer: Restore with deionized water (or equivalent)

Concentration: 5.0 mg/mL

Buffer:

Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2

Stabilizer: None

Preservative: 0.01 % (w/v) Sodium Azide

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: 4 °C,-20 °C

Storage Comment: Store vial at 4° C prior to restoration.   For extended storage aliquot contents and freeze at -20° C or below.  Avoid cycles of freezing and thawing.  Centrifuge product if not completely clear after standing at room temperature.  This product is stable for several weeks at 4° C as an undiluted liquid.  Dilute only prior to immediate use. 

Expiry Date: 12 months

References for anti-ISG15 antibody (ABIN103335)

Gall, Pryke, Abraham, Mizuno, Botto, Sali, Broeckel, Haese, Nilsen, Placzek, Morrison, Heise, Streblow, DeFilippis: "Emerging Alphaviruses Are Sensitive to Cellular States Induced by a Novel Small-Molecule Agonist of the STING Pathway." in: Journal of virology, Vol. 92, Issue 6, (2018) (PubMed).

Ezzati, Komher, Severini, Coombs: "Comparative proteomic analyses demonstrate enhanced interferon and STAT-1 activation in reovirus T3D-infected HeLa cells." in: Frontiers in cellular and infection microbiology, Vol. 5, pp. 30, (2015) (PubMed).

Fields, Dumaop, Adame, Ellis, Letendre, Grant, Masliah: "Alterations in the levels of vesicular trafficking proteins involved in HIV replication in the brains and CSF of patients with HIV-associated neurocognitive disorders." in: Journal of neuroimmune pharmacology : the official journal of the Society on NeuroImmune Pharmacology, Vol. 8, Issue 5, pp. 1197-209, (2014) (PubMed).

Coombs: "HeLa cell response proteome alterations induced by mammalian reovirus T3D infection." in: Virology journal, Vol. 10, pp. 202, (2013) (PubMed).

Berard, Cortens, Krokhin, Wilkins, Severini, Coombs: "Quantification of the host response proteome after mammalian reovirus T1L infection." in: PLoS ONE, Vol. 7, Issue 12, pp. e51939, (2013) (PubMed).

Loch, Cuccherini, Leach, Strickler: "Deubiquitylase, deSUMOylase, and deISGylase activity microarrays for assay of substrate preference and functional modifiers." in: Molecular & cellular proteomics : MCP, Vol. 10, Issue 1, pp. M110.002402, (2011) (PubMed).

Target Details for ISG15

Target: ISG15 (ISG15 Ubiquitin-Like Modifier (ISG15))

Alternative Name: ISG15 (ISG15 Products)

Background: Background: Ubiquitin-like proteins fall into two classes: the first class, ubiquitin-like modifiers (UBLs) function as modifiers in a manner analogous to that of ubiquitin.  Examples of UBLs are SUMO, Rub1 (also called Nedd8), Apg8 and Apg12.  Proteins of the second class include parkin, RAD23 and DSK2, are designated ubiquitin-domain proteins (UDPs).  These proteins contain domains that are related to ubiquitin but are otherwise unrelated to each other.  In contrast to UBLs, UDPs are not conjugated to other proteins. ISG15 (Interferon Stimulating Gene-15) shows no amino acid sequence homology to cytokines and is synthesized as a precursor that is activated through processing by a thiol protease.   ISG15 is secreted by monocytes and lymphocytes. Synthesis is induced in response to IFN-a or IFN-b or IFN-, but not IFN-g.  ISG15 expression is induced also by overexpression of some interferon regulatory factors that have been shown to play a role in the transcriptional regulation of IFN genes.  ISG15 is secreted also by cell lines of monocyte (U937 cell line), T-lymphocyte, B-lymphocyte (DAUDI cells), human fibroblasts, and epithelial origins. The induction of terminal differentiation in human melanoma cells is associated, among other things, with alterations in the expression of ISG15.   Intracellularly ISG15 has been shown to function as a ubiquitin homologue. It is known also as UCRP (ubiquitin cross-reactive protein). Serpin 2a (spi2a), a member of the serine protease inhibitor (serpin) protein family that is highly induced in macrophages during bacillus Calmette-Guerin infection has been shown to bind ISG15.  ISG15 has been shown to modulate immune cell function. It possesses activities of  cytokines and induces production of IFN-g.  It enhances proliferation and functions of natural killer and LAK cells.

Gene ID: 9636, 4826774

UniProt: P05161