For ResearchUseOnly. NotForUseInDiagnostic Procedures. ® 2014 Cell Signaling Technology, Inc. Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc. Store at -20°C protocols. See www.cellsignal.com forindividualcomponentapplications,dilutionsandadditionalapplication Dg—dog Species Cross-Reactivity Key: Applications Key: towards theplanarcellpolaritypathway(11-12). by bindingtoDshproteinsanddirectingactivity (4,10). NakedinhibitsthecanonicalWnt/ thase kinase-3β,therebypromoting-cateninstabilization (Dsh) proteins(Dvl1,Dvl2andDvl3)inhibitglycogensyn subsequent degradationofβ-catenin(8-9).TheDisheveled and and promotestheGSK-3β-mediatedphosphorylation Axin1 complexeswithAPC,GSK-3β,Dvland-catenin proteins Axin1andAxin2negativelyregulateWntsignaling. the absenceofWntstimulation,multidomainscaffold and subsequentlyrecruitsAxintothemembrane(5-7).In by kinasessuchasGSK-3andCK1 is phosphorylated as LRP5andLRP6(4).UponstimulationwithWnt, the low-densitylipoproteinreceptor(LDLR)family, such pathway requiresacoreceptorfromproteinsbelongingto signaling pathways(3).ThecanonicalWnt/ bind totheFrizzledfamilyofproteinsandactivateseveral is involvedinseveraltypesofcancers(2).Wntmembers ment (1).AberrantactivationofWntsignalingpathways glycoproteins thatplayimportantrolesinanimaldevelop Background: TheWntfamilyincludesseveralsecreted antibody.primary antibody toperformtwowesternblotexperimentswitheach within theWntsignalingpathway. Thekitincludesenough provides aneconomicalmeansofdetectingintegralproteins Description: TheWntSignalingAntibodySamplerKit Anti-rabbit IgG,HRP-linkedAntibody Axin1 (C76H11)RabbitmAb Naked2 (C67C4)RabbitmAb Naked1 (C30F10)RabbitmAb Dvl3 Antibody Dvl2 (30D2)RabbitmAb Phospho-LRP6 (Ser1490)Antibody LRP6 (C47E12)RabbitmAb Wnt5a/b (C27E8)RabbitmAb Wnt3a (C64F2)RabbitmAb Products Included #2915 n Antibody SamplerKit Wnt Signaling 3 Pg—pig  (9 x20µl) 1 Kit Sc—S. cerevisiae W—Western H—human IP—Immunoprecipitation All—all species expected β-catenin pathway M—mouse β-catenin R—rat Product # IHC—Immunohistochemistry - 7074 2087 2073 2201 3218 3224 2568 3395 2530 2721 - Species enclosed inparenthesesarepredicted toreactbasedon100% homology. Hm—hamster peptide affinitychromotagraphy. Dvl3. PolyclonalantibodiesarepurifiedbyProteinAand to residuessurroundingthecarboxyterminusofhuman immunizing animalswithasyntheticpeptidecorresponding human LRP6.Thetotalpolyclonalantibodyisproducedby peptide correspondingtoresiduessurroundingSer1490of duced byimmunizinganimalswithasyntheticphospho Axin1. Thephospho-specificpolyclonalantibodyispro produced byimmunizinganimalswithrecombinanthuman human Naked2.ThemonoclonalantibodyforAxin1is of humanDvl2,Leu53Naked1,andAla217 Glu176 ofhumanWnt5a,Met988LRP6,Gly176 ing toresiduessurroundingLeu256ofhumanWnt3a, by immunizinganimalswithasyntheticpeptidecorrespond Source/Purification: Monoclonalantibodyisproduced enous levelsoftheirrespectivetotalproteins. antibodies intheWntSignalingSamplerKitdetectendog atSer1490.Theremaining protein whenphosphorylated (Ser1490) AntibodydetectsendogenouslevelsofLRP6 conditioned mediumfromL/Wnt3acells.Phospho-LRP6 detects transfectedWnt3aproteininLcellsandthe Specificity/Sensitivity: Wnt3a(C64F2)RabbitmAb Quantity 100 µl 20 µl 20 µl 20 µl 20 µl 20 µl 20 µl 20 µl 20 µl 20 µl Mk—monkey ChIP—Chromatin Immunoprecipitation 180, 210kDa 180, 210kDa Mi—mink 88–93 kDa 90–95 kDa 59, 61kDa 59, 61kDa Mol. Wt. rev. 06/16 110 kDa 45 kDa 42 kDa C—chicken Rabbit IgG Rabbit IgG Rabbit IgG Rabbit IgG Rabbit IgG Rabbit IgG Rabbit IgG Rabbit IgG Rabbit IgG Isotype Goat Dm—D. melanogaster IF—Immunofluorescence - - - - U.S. PatentNo.5,675,063 products. and acompletelistingofrecommendedcompanion Please visitwww.cellsignal.com forvalidateddata Western blotting Recommended AntibodyDilutions: aliquot theantibodies. NaCl, 100µg/mlBSAand50%glycerol.Storeat–20°C. Storage: Suppliedin10mMsodiumHEPES(pH7.5),150 (12)  (11)  (10)  Background References: (9)  (8)  (7)  (6)  (5)  (4)  (3)  (2)  (1)  Salahshor, S.andWoodgett, J.R.(2005)JClinPathol58, Luo, W. andLin,S.C.Neurosignals13,99–113. Davidson, G.etal.(2005)Nature438,867–72. Zeng, X.etal.(2005)Nature438,873–7. Tamai, K.etal.(2000)Nature407,530–5. Logan, C.Y. andNusse,R.(2004)AnnuRevCellDevBiol Kohn, A.D.andMoon,R.T. CellCalcium38,439–46. Giles, R.H.etal.(2003)BiochimBiophysActa1653,1–24. Cadigan, K.M.andNusse,R.(1997)GenesDev11, Yan, D.etal.(2001)ProcNatlAcadSciUSA98,3802–7. Wharton, K.A.etal.(2001)DevBiol234,93–106. Boutros, M.andMlodzik,(1999)MechDev83,27–37. 225–36. 20, 781–810. 3286–305. X—Xenopus Support Orders Web F—Flow cytometry Z—zebra fish n n n www.cellsignal.com [email protected] 877-678-TECH (8324) [email protected] 877-616-CELL (2355) E-P—ELISA-Peptide B—bovine 1:1000 Do not