For ResearchUseOnly. NotForUseInDiagnostic Procedures. © 2013 Cell Signaling Technology, Inc. XP® and Cell Signaling Technology® are trademarks of Cell Signaling Technology, Inc. Store at -20ºC Dg—dog Species Cross-Reactivity Key: Applications Key: Background References: . ing toresiduessurroundingLeu248ofhumanRab11FIP1 by immunizinganimalswithasyntheticpeptidecorrespond Source/Purification: Monoclonalantibodyisproduced recognizes endogenouslevelsoftotalRab11FIP1protein. Specificity/Sensitivity: Rab11FIP1(D9D8P)RabbitmAb effectors incarcinogenesis(8,9). have implicatedRab11familyproteinsandtheirinteracting that bindArf6andRab11proteins(5-7).Researchstudies Rab11FIP3 andRab11FIP4belongtotheclassIIsubfamily are categorizedasclassIsubfamilymembers,whereas domain structures,Rab11FIP1,Rab11FIP2andRab11FIP5 have beenidentified.Basedonsequencehomologyand ing domain(3,4).AtleastsixmembersoftheRab11FIPs carboxy-terminalRab11-bind- (Rab11FIPs) viaaconserved interactwiththeRab11familyinteracting membrane traffic(1,2).TheGTP-boundactivestateRab11 thought toplayanessentialroleinregulatingendocytic lated Rab11a,Rab11b,andRab25.TheyaresmallGTPases Background: Rab11familyproteinsconsistofcloselyre (9)  (8)  (7)  (6)  (5)  (4)  (3)  (2)  (1)  Rabbit mAb#8457. using Rab11FIP1(D9D8P)RabbitmAband Western blotanalysisofextractsfromMCF7andSNB19cells #12849 milk, 1XTBS,0.1% Tween-20 at4°Cwithgentle shaking,overnight. IMPORTANT: Forwesternblots,incubatemembrane withdilutedantibodyin5%w/vnonfatdry Muller, P.A. etal.(2009)Cell139,1327-41. Zhang, J.etal.(2009)JClinInvest119,2171-83. Hickson, G.R.etal.(2003)MolBiolCell14,2908-20. Lindsay, A.J.andMcCaffrey, M.W. (2004)JCellSci Meyers, J.M.andPrekeris,R.(2002)JBiolChem277, Prekeris, R.etal.(2001)JBiolChem276,38966-70. Hales, C.M.etal.(2001)JBiolChem276,39067-75. Ullrich, O.etal.(1996)JCellBiol135,913-24. Bock, J.B.etal.(2001)Nature409,839-41. 117, 4365-75. 49003-10. n Rab11FIP1 (D9D8P) RabbitmAb 3 Endogenous Applications W, IP, IF-IC Pg—pig  (10 westernblots) 100 µl Sc—S. cerevisiae W—Western H—human Species Cross-Reactivity* IP—Immunoprecipitation Ce—C. elegans H, M,Mk M—mouse β -Actin (D6A8)

Hr—horse R—rat IHC—Immunohistochemistry - - Hm—hamster cells usingRab11FIP1(D9D8P)RabbitmAb. Western blotanalysisofextractsfromHeLa,HepG2,andBaF3 Molecular Wt. 85 kDa 100 140 200 kDa 100 140 200 20 30 40 50 80 kDa All—all species expected 20 30 40 50 60 80 Mk—monkey ChIP—Chromatin Immunoprecipitation

MCF7 HeLa SNB19 Mi—mink Rabbit IgG**

Hep G2 New 07/13 Isotype β-Actin Rab11FIP1 BaF3 Species enclosed inparenthesesarepredicted toreactbasedon100% homology. Rab11FIP1 C—chicken Dm—D. melanogaster IF—Immunofluorescence UniProt ID#Q6WKZ4 GeneID#80223 of recommended complementary products. of recommendedcomplementary Please visitwww.cellsignal.com foracompletelisting for thisproductatwww.cellsignal.com. For productspecificprotocolspleaseseethewebpage Immunofluorescence (IF-IC) 1:100 Immunoprecipitation 1:100 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 DRAQ5 (fluorescent DNAdye). Rabbit mAb(green).Bluepseudocolor=DRAQ5 and SNB19(negative,lower)cellsusingRAB11FIP1 (D9D8P) Confocal immunofluorescentanalysisofMCF7 (positive,upper) detect thisantibody. antibodiesmustbeusedto Anti-rabbit secondary X—Xenopus SNB19 MCF7 ® Support isaregisteredtrademark ofBiostatusLimited. Orders Web F—Flow cytometry Z—zebrafish n n n www.cellsignal.com [email protected] 877-678-TECH (8324) [email protected] 877-616-CELL (2355) E-P—ELISA-Peptide B—bovine ® #4084 1:1000 kDa 1 2 3

200 140 100 80 Rab11FIP1

60 IgG heavy chain 50 40 30

20

10 – + – Rabbit (DA1E) mAb IgG XP® Isotype Control – – + Rab11FIP1 (D9D8P) Rabbit mAb

Immunoprecipitation of Rab11FIP1 from HeLa cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Rab11FIP1 (D9D8P) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Rab11FIP1 (D9D8P) Rabbit mAb.

© 2013 Cell Signaling Technology, Inc. © 2013 Cell Signaling Technology, Orders n 877-616-CELL (2355) [email protected] Support n 877-678-TECH (8324) [email protected] Web n www.cellsignal.com