Ubiquitin-Like Modifier (ISG15) antibody

Antigen: Ubiquitin-Like Modifier (ISG15)

Synonyms: G1P2, UCRP, IFI15, G1p2, IP17, Irfp, IGI15, MGC18616, MGC103144, MGC130321

Clonality: Polyclonal

Host: Rabbit

Reactivity: Mouse (Murine), Human, Rat (Rattus)

Application: ELISA, Western Blotting (WB), Immunoprecipitation (IP)

Catalog no.: ABIN103335

Additional Information

Alternative name: ISG15

Immunogen: This purified antibody was prepared from rabbitserum after repeated immunizations with recombinant humanISG15 protein.

Format: Liquid (sterile filtered)

Description: Ubiquitin-like proteins fall into twoclasses: the first class, ubiquitin-like modifiers (UBLs) function asmodifiers in a manner analogous to that of ubiquitin. Examplesof 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). Theseproteins contain domains that are related to ubiquitin but areotherwise unrelated to each other. In contrast to UBLs, UDPsare not conjugated to other proteins. ISG15 (InterferonStimulating Gene-15) shows no amino acid sequence homologyto cytokines and is synthesized as a precursor that is activatedthrough processing by a thiol protease. ISG15 is secreted bymonocytes and lymphocytes. Synthesis is induced in response toIFN-alpha or IFN- beta or IFN- ∞ , but not IFN- gamma . ISG15 expression is induced also by overexpression of some interferon regulatoryfactors that have been shown to play a role in the transcriptionalregulation of IFN genes. ISG15 is secreted also by cell lines ofmonocyte (U937 cell line), T-lymphocyte, B-lymphocyte (DAUDIcells), human fibroblasts, and epithelial origins. The induction ofterminal 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-reactiveprotein). Serpin 2a (spi2a), a member of the serine protease inhibitor (serpin) protein family that is highly induced inmacrophages during bacillus Calmette-Guerin infection has been shown to bind ISG15. ISG15 has been shown to modulateimmune cell function. It possesses activities of cytokines and induces production of IFN- gamma . It enhances proliferation and functions of natural killer and LAK cells.

Specificity: This product is an IgG fraction antibody purified from monospecific antiserum by a multi-step processwhich includes delipidation, salt fractionation and ion exchange chromatography followed by extensive dialysis against thebuffer stated above. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Rabbit Serum.

Application Details

Application Notes: Recommended Dilutions: ELISA 1:2,000 - 1:10,000 WESTERN BLOT 1:200 - 1:1,000 IMMUNOHISTOCHEMISTRY User Optimized OTHER APPLICATIONS User Optimized. 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). Other intrinsic bands are readily detectable at lower dilutions. An 18.5 kDa band corresponding to human ISG15 is detected. Most human cell lysates can be used as apositive control without induction or stimulation.

Concentration: 5.0 mg/ml (by UV absorbance at 280 nm)

Buffer: Stabilizer: None. Preservative: 0.01% (w/v) Sodium Azide. Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2.

Storage: Store vial at 4° C prior to restoration. Restore with 0.1 ml of deionized water (or equivalent). Forextended storage aliquot contents and freeze at -20° C or below. Avoid cycles of freezing and thawing. Centrifuge product ifnot completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undilutedliquid. Dilute only prior to immediate use. Expiration date is one (1) year from date of restoration.

Research Area: Proteolysis / Ubiquitin, Hormones, Cytokines

Restrictions: For Research Use only

Images

Immunoblot of hISG15 fusion protein. Anti-hISG15 antibody, generated by immunization with recombinant human ISG15, was tested by immunoblot against a hISG15-GFP fusion protein produced in E.coli cell lysate soluble fraction. Dilution of the antibody between 1:200 and 1:1,000 showed strong reactivity specifically with hISG15 and ISG15 coupled proteins. Free hISG15 is indicated by the arrowhead. In this blot the antibody was used at a 1:200 dilution incubated overnight at 4