Revision 3

#4813Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

34, 39

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q9UMW8

Entrez-Gene Id:

11274

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

USP18 (D4E7) Rabbit mAb detects endogenous levels of total USP18 protein. The doublet band detected by western blot represents full length (39 kDa) and amino-terminal deleted derivative of USP18 (8).

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro45 of human USP18 protein.

Background

Ubiquitinating enzymes (UBEs) catalyze protein ubiquitination, a reversible process countered by deubiquitinating enzyme (DUB) action. Five DUB subfamilies are recognized, including the USP, UCH, OTU, MJD, and JAMM enzymes (1,2). USP18 (also known as UBP43) is a deubiquitinase best known for catalyzing the removal of ISG15, an interferon-regulated ubiquitin-like protein, from conjugated proteins (3). Removal of ISG15 from target proteins by the USP18 peptidase maintains the critical cellular balance of ISG15-conjugated proteins important for normal development and brain function (4,5). Following induction by IFN or LPS (6), USP18 binds the INF receptor subunit IFNAR2 and inhibits signal transduction through the Jak/Stat pathway (7). USP18 regulation of IFN signaling inhibits IFN-mediated apoptosis and does not necessarily rely on USP18 peptidase activity (8). As the therapeutic use of recombinant IFN can lead to refractory IFN signaling and a less effective response, the combination of IFN treatment and regulation of USP18 expression may produce a more positive outcome (9).

  1. Nijman, S.M. et al. (2005) Cell 123, 773-86.
  2. Nalepa, G. et al. (2006) Nat Rev Drug Discov 5, 596-613.
  3. Malakhov, M.P. et al. (2002) J Biol Chem 277, 9976-81.
  4. Rempel, L.A. et al. (2007) Reprod Biol Endocrinol 5, 13.
  5. Ritchie, K.J. et al. (2002) Genes Dev 16, 2207-12.
  6. Malakhova, O. et al. (2002) J Biol Chem 277, 14703-11.
  7. Malakhova, O.A. et al. (2006) EMBO J 25, 2358-67.
  8. Potu, H. et al. (2010) Cancer Res 70, 655-65.
  9. Sarasin-Filipowicz, M. et al. (2009) Mol Cell Biol 29, 4841-51.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 3
#4813

USP18 (D4E7) Rabbit mAb

Western Blotting Image 1: USP18 (D4E7) Rabbit mAb Expand Image
Western blot analysis of extracts from THP-1 cells, untreated or LPS-treated (1 µg/ml, 24 hours), using USP18 (D4E7) Rabbit mAb.
Immunoprecipitation Image 1: USP18 (D4E7) Rabbit mAb Expand Image
Immunoprecipitation of USP18 from THP-1 extracts treated with TPA (12-O-Tetradecanoylphorbol-13-Acetate) #4174 (80nM overnight) and Lipopolysaccharides (LPS) #14011 (1μg/mL overnight). Lane 1 is 10% input, lane 2 is USP18 (D4E7) Rabbit mAb, and lane 3 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900. Western blot analysis was perfomed using USP18 (D4E7) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.