USP8 (D18F6) Rabbit mAb www.cellsignal.com - 20ºC Store at Support: 877-678-TECH (8324) [email protected]

100 µl (10 western blots) Orders: 877-616-CELL (2355) [email protected]

Entrez-Gene ID #9101

#11832 rev. 06/09/14 UniProt ID #P40818

For Research Use Only. Not For Use In Diagnostic Procedures.

Applications Species Cross-Reactivity* Molecular Wt. Isotype Storage: Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% W, IP H, Mk 140 kDa Rabbit IgG** Endogenous sodium azide. Store at –20°C. Do not aliquot the antibody. *Species cross-reactivity is determined by western blot. **Anti-rabbit secondary antibodies must be used to Background: Ubiquitinating enzymes (UBEs) catalyze kDa detect this antibody. ubiquitination, a reversible process countered by deubiquiti- nating enzyme (DUB) action (1,2). Five DUB subfamilies are 200 Recommended Antibody Dilutions: recognized, including the USP, UCH, OTU, MJD, and JAMM Western blotting 1:1000 enzymes. The deubiquitinating enzyme ubiquitin-specific USP8 (Myc/DDK-tagged) Immunoprecipitation 1:100 protease 8 (USP8/UBPy) is a cysteine protease belonging to the 140 USP8 (Endogenous) Background References: USP/UBP subfamily. Research studies have shown that USP8 is an essential growth-regulated enzyme indespensible for cell (1) Nijman, S.M. et al. (2005) Cell 123, 773-86. 100 proliferation and survival (3,4). Indeed, conditional knock-out of (2) Nalepa, G. et al. (2006) Nat Rev Drug Discov 5, 596-613. murine USP8 was shown to promote a dramatic loss in expres- (3) Naviglio, S. et al. (1998) EMBO J 17, 3241-50. sion of receptor tyrosine kinases, including EGFR, ErbB3, and 80 c-Met (4). In agreement with these findings, USP8 inactivation – + hUSP8-Myc/DDK (4) Niendorf, S. et al. (2007) Mol Cell Biol 27, 5029-39. leads to enhanced ubiquitination of ligand-activated EGFR (5,6). Western blot analysis of extracts from 293T cells, either mock (5) Alwan, H.A. and van Leeuwen, J.E. (2007) J Biol Chem Furthermore, phosphorylation of USP8 at Ser680 results in its 282, 1658-69. binding of 14-3-3, catalytic inactivation, and reduced EGFR transfected (-) or transfected with a construct expressing Myc/DDK- tagged full-length human USP8 (hUSP8-Myc/DDK +), using USP8 deubiquitination (7). It appears as though USP8, in conjunction (6) Mizuno, E. et al. (2005) Mol Biol Cell 16, 5163-74. (D18F6) Rabbit mAb. with components of the ESCRT-0 complex, plays an integral (7) Mizuno, E. et al. (2007) Exp Cell Res 313, 3624-34. role in the early endosomal sorting machinery that functions to (8) Row, P.E. et al. (2006) J Biol Chem 281, 12618-24. protect EGFR from lysosomal degradation (8,9). (9) Berlin, I. et al. (2010) J Biol Chem 285, 34909-21. Specificity/Sensitivity: USP8 (D18F6) Rabbit mAb recog- nizes endogenous levels of total USP8 protein. Source/Purification: Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu614 of human USP8 protein.

kDa 293T MCF7 U-2 OS NCI-H460DLD-1 A549 U251 KARPAS-299A-431 A-375 COS-7 Western blot analysis of extracts from various cell lines 200 using USP8 (D18F6) Rabbit mAb.

140 USP8

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Thank you for your recent purchase. If you would like to provide a review visit www.cellsignal.com/comments. IMPORTANT: For western blots, incubate membrane with diluted antibody in 5% w/v BSA, 1X TBS, 0.1% Tween®20 at 4°C with gentle shaking, overnight.

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Applications: W—Western IP—Immunoprecipitation IHC—Immunohistochemistry ChIP—Chromatin Immunoprecipitation IF—Immunofluorescence F—Flow cytometry E-P—ELISA-Peptide Species Cross-Reactivity: H—human M—mouse R—rat Hm—hamster Mk—monkey 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 All—all species expected Species enclosed in parentheses are predicted to react based on 100% homology.