ISG15/UCRP Antibody (851701) [Unconjugated]

A549 cells transfected with HA-ISG15 were treated briefly with IFN gamma or carrier only. 500 µg of cell lysates were subjected to immunoprecipitation (IP) with 20 µg A-830 using Protein G Sepharose. IP’d material was resolved on reducing SDS-PAGE gels and Western Blots were developed using PVDF membranes and anti-HA epitope (R&D Systems MAB060) at 0.5 µg/ml followed by HRP-labeled anti-mouse (R&D Systems # HAF007) secondary antibody at 1:2000 dilution. Free (unconjugated) HA-ISG15 was visualized in both treated and untreated cells, while ISGylated proteins were clearly recovered (IP’d) from the interferon-treated cell lysates.

Product Discontinued

ISG15/UCRP Antibody (851701) [Unconjugated] Summary

• Additional Information: Product is in the process of being discontinued.
• Immunogen: Full-length recombinant human ISG15/UCRP
  Accession # P05161
• Specificity: This antibody will recover ISGylated proteins in immunoprecipitation applications. This antibody detects ISGylated proteins in Western blots.
• Source: N/A
• Isotype: IgG2a
• Clonality: Monoclonal
• Host: Rat
• Gene: ISG15
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunoprecipitation 10-20 ug/mL
  • Western Blot 0.1-0.5 ug/mL

Packaging, Storage & Formulations

• Storage: Store the unopened product at -20 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Storage below -20 °C is not recommended. Do not use past expiration date.
• Buffer: Supplied as a solution in PBS containing Glycerol and Sodium Azide.
• Preservative: Sodium Azide

Notes

* Contains <0.1% Sodium Azide, which is not hazardous at this concentration according to GHS classifications. Refer to SDS for additional information and handling instructions.

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for ISG15/UCRP Antibody (851701) [Unconjugated]

• G1P2
• G1P2interferon-induced 17-kDa/15-kDa protein
• IFI15
• IFI15hUCRP
• interferon, alpha-inducible protein (clone IFI-15K)
• Interferon-induced 15 kDa protein
• Interferon-induced 17 kDa protein
• interferon-stimulated protein, 15 kDa
• IP17
• ISG15 ubiquitin-like modifier
• ISG15
• Ubiquitin cross-reactive protein
• ubiquitin-like protein ISG15
• UCRP
• UCRPIP17

Background

Interferon­stimulated Gene 15 (ISG15), also known as Ubiquitin Cross­reacting Protein (UCRP), is a Ubiquitin­like protein that is covalently coupled to target proteins in a process termed ISGylation. It is a 165 amino acid (aa) polypeptide with a predicted molecular weight of 18 kDa. ISG15/UCRP exhibits 66% aa sequence identity with its mouse ortholog. Structurally, ISG15/UCRP consists of two tandem Ubiquitin­like domains that share a similar 3­dimensional structure with Ubiquitin and other Ubiquitin­like modifiers including NEDD8 and SUMO1. Modification of targets by ISG15/UCRP occurs in a stepwise enzymatic process similar to that of Ubiquitin. Enzymes regulating ISGylation include the activating (E1) enzyme UBE1L, the conjugating (E2) enzyme UbcH8, and ligases (E3) such as HERC5. Removal of ISG15/UCRP is catalyzed by the deconjugating enzyme UBP43/USP18. Functionally, ISG15/UCRP has putative roles in the immune response and tumorigenesis. This is reflected by intracellular ISG15/UCRP targets that include Cyclin D1, tumor suppressor p63, IRF3, and a range of viral proteins. It is induced by type 1 interferons and microbial infection, and knockout mice exhibit an increased sensitivity to infection by some viruses. ISG15/UCRP can also be secreted by cells of the immune system and may act in a cytokine­like manner. For instance, it is produced by human granulocytes in response to mycobacterium exposure, and natural killer cells and T cells respond to extracellular ISG15/UCRP with IFN­gamma production. Further supporting a role in immune function, ISG15/UCRP mutations are associated with MSMD, an inherited disorder characterized by increased susceptibility to mycobacterial infection.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for ISG15/UCRP Antibody (A-830)(2)

Publications using A-830

• A Pinto-Fern, M Salio, T Partridge, J Chen, G Vere, H Greenwood, CS Olie, A Damianou, HC Scott, HJ Pegg, A Chiarenza, L Díaz-Saez, P Smith, C Gonzalez-L, B Patel, E Anderton, N Jones, TR Hammonds, K Huber, R Muschel, P Borrow, V Cerundolo, BM Kessler Deletion of the deISGylating enzyme USP18 enhances tumour cell antigenicity and radiosensitivity Br J Cancer, 2020-11-20;0(0):. 2020-11-20 [PMID: 33214684] (Immunoprecipitation, Human)
• L Yoo, AR Yoon, CO Yun, KC Chung Covalent ISG15 conjugation to CHIP promotes its ubiquitin E3 ligase activity and inhibits lung cancer cell growth in response to type I interferon Cell Death Dis, 2018-01-24;9(2):97. 2018-01-24 [PMID: 29367604] (Western Blot, Human)

Bioinformatics

• Gene Symbol: ISG15
• Uniprot:
  • P05161(Human)