For ResearchUseOnly. NotForUseInDiagnostic Procedures. © 2012 Cell Signaling Technology, Inc. XP® and Cell Signaling Technology® are trademarks of Cell Signaling Technology, Inc. Store at -20°C Dg—dog Species Cross-Reactivity Key: Applications Key: Western blotanalysisofextracts fromvariouscelllinesusingADRM1(D9Z1U)RabbitmAb. . ing toresiduesneartheaminoterminusofhumanADRM1 by immunizinganimalswithasyntheticpeptidecorrespond Source/Purification: Monoclonalantibodyisproduced recognizes endogenouslevelsoftotalADRM1protein. Specificity/Sensitivity: ADRM1(D9Z1U)RabbitmAb in mammaliandevelopment(6). thus highlightingaphysiologicroleforendogenousADRM1 murine Adrm1knockoutresultsindefectivegametogenesis, a mechanismtoaccelerateubiquitinchaindisassembly. A as tidase activityofUCHL5/UCH37(3-5),perhapsserving tobindandenhancetheisopep- mammalian ADRM1serves ubiquitin (Pru)domain.Thecarboxy-terminaldomainof homology domainknownasthepleckstrin-likereceptorfor the (1,2)thatbindsubiquitinviaapleckstrin Rad23, Dsk2,andDdi1.ADRM1isaubiquitinreceptorof lating molecule1),andthreefamiliesofshuttlingfactors, Rpn10/S5a/PSMD4 andRpn13/ADRM1(Adhesion-regu- associated withtheproteasome:twoproteasomesubunits, Background: Currently, therearefiveubiquitinreceptors #12019 milk, 1XTBS,0.1% Tween-20 at4°Cwithgentle shaking,overnight. IMPORTANT: Forwesternblots,incubatemembrane withdilutedantibodyin5%w/vnonfatdry 100 140 200 kDa 20 30 40 50 60 80 n ADRM1 (D9Z1U)RabbitmAb 3 Endogenous Applications Pg—pig  (10 westernblots) 100 µl W, IP

MCF7 Sc—S. cerevisiae W—Western PC-3

U-2 OS H—human SW620 Species Cross-Reactivity* IP—Immunoprecipitation Ce—C. elegans U-251 MG H, M,R,Mk

M—mouse MOLT-4

A20 Hr—horse Raw 264.7 R—rat

IHC—Immunohistochemistry H-4-II-E - COS-7 Hm—hamster +), usingADRM1(D9Z1U)RabbitmAb. DDK-tagged full-lengthhumanADRM1(hADRM1-Myc/DDK; transfected (-)orwithaconstructexpressingMyc/ Western blotanalysisofextractsfrom293Tcells,mock SK-MEL-2 Molecular Wt. 100 140 200 kDa 20 30 40 50 60 80 ADRM1 45 kDa All—all species expected Mk—monkey – ChIP—Chromatin Immunoprecipitation + hADRM1-Myc/DDK ADRM1 (Endogenous) ADRM1 (Myc/DDK-tagged) Mi—mink Rabbit IgG** New 10/12 Isotype Species enclosed inparenthesesarepredicted toreactbasedon100% homology. C—chicken Dm—D. melanogaster IF—Immunofluorescence Swiss-Prot Acc.#Q16186 - ID#11047 Rabbit (DA1E)mAbIgGXP Immunoprecipitation ofADRM1from293Tcell extractsusing Rabbit mAb. Western blotdetectionwasperformedusingADRM1(D9Z1U) ADRM1 (D9Z1U)RabbitmAb(lane3).Lane1 is10%input. Background References: products. of recommendedcomplementary Please visitwww.cellsignal.com foracompletelisting for thisproductatwww.cellsignal.com. For productspecificprotocolspleaseseethewebpage Immunoprecipitation 1:50 Western blotting Recommended AntibodyDilutions: ** *Species cross-reactivityisdeterminedbywesternblot. sodium azide.Storeat–20°C.Donotaliquottheantibody. mM NaCl,100µg/mlBSA,50%glycerolandlessthan0.02% Storage: Suppliedin10mMsodiumHEPES(pH7.5),150 (6)  (5)  (4)  (3)  (2)  (1)  detect thisantibody. antibodiesmustbeusedto Anti-rabbit secondary Al-Shami, A.etal.(2010)PLoSOne5,e13654. Qiu, X.B.etal.(2006)EMBOJ25,5742-53. Hamazaki, J.etal.(2006)EMBOJ25,4524-36. Yao, T. etal.(2006)NatCellBiol8,994-1002. Husnjak, K.etal.(2008)Nature453,481-8. Schreiner, P. etal.(2008)Nature453,548-52. X—Xenopus Support 100 140 200 kDa Orders 10 20 30 40 50 60 80 Web F—Flow cytometry 1 – – Z—zebrafish n n n + – 2 www.cellsignal.com [email protected] 877-678-TECH (8324) [email protected] 877-616-CELL (2355) ® – + 3 IsotypeControl#3900(lane2)or ADRM1 (D9Z1U)RabbitmAb XP Rabbit (DA1E)mAbIgG E-P—ELISA-Peptide ADRM1 IgG LightChain ® B—bovine Isotype Control 1:1000