Wnt Signaling Antibody Sampler Kit Store at -20°C 3 n 1 Kit Orders n 877-616-CELL (2355) (9 x 20 µl) [email protected] Support n 877-678-TECH (8324) [email protected] Web n www.cellsignal.com rev. 06/16 #2915 For Research Use Only. Not For Use In Diagnostic Procedures. Products Included Product # Quantity Mol. Wt. Isotype Storage: Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM Wnt3a (C64F2) Rabbit mAb 2721 20 µl 42 kDa Rabbit IgG NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibodies. Wnt5a/b (C27E8) Rabbit mAb 2530 20 µl 45 kDa Rabbit IgG Recommended Antibody Dilutions: LRP6 (C47E12) Rabbit mAb 3395 20 µl 180, 210 kDa Rabbit IgG Western blotting 1:1000 Phospho-LRP6 (Ser1490) Antibody 2568 20 µl 180, 210 kDa Rabbit IgG Please visit www.cellsignal.com for validated data and a complete listing of recommended companion Dvl2 (30D2) Rabbit mAb 3224 20 µl 90–95 kDa Rabbit IgG products. Dvl3 Antibody 3218 20 µl 88–93 kDa Rabbit IgG U.S. Patent No. 5,675,063 Naked1 (C30F10) Rabbit mAb 2201 20 µl 59, 61 kDa Rabbit IgG Naked2 (C67C4) Rabbit mAb 2073 20 µl 59, 61 kDa Rabbit IgG Axin1 (C76H11) Rabbit mAb 2087 20 µl 110 kDa Rabbit IgG Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat See www.cellsignal.com for individual component applications, dilutions and additional application protocols. Description: The Wnt Signaling Antibody Sampler Kit Specificity/Sensitivity: Wnt3a (C64F2) Rabbit mAb Background References: provides an economical means of detecting integral proteins detects transfected Wnt3a protein in L cells and the (1) Cadigan, K.M. and Nusse, R. (1997) Genes Dev 11, within the Wnt signaling pathway. The kit includes enough conditioned medium from L/Wnt3a cells. Phospho-LRP6 3286–305. antibody to perform two western blot experiments with each (Ser1490) Antibody detects endogenous levels of LRP6 primary antibody. protein when phosphorylated at Ser1490. The remaining (2) Giles, R.H. et al. (2003) Biochim Biophys Acta 1653, 1–24. antibodies in the Wnt Signaling Sampler Kit detect endog- (3) Kohn, A.D. and Moon, R.T. Cell Calcium 38, 439–46. Background: The Wnt family includes several secreted enous levels of their respective total proteins. glycoproteins that play important roles in animal develop- (4) Logan, C.Y. and Nusse, R. (2004) Annu Rev Cell Dev Biol ment (1). Aberrant activation of Wnt signaling pathways Source/Purification: Monoclonal antibody is produced 20, 781–810. is involved in several types of cancers (2). Wnt members by immunizing animals with a synthetic peptide correspond- (5) Tamai, K. et al. (2000) Nature 407, 530–5. bind to the Frizzled family of proteins and activate several ing to residues surrounding Leu256 of human Wnt3a, signaling pathways (3). The canonical Wnt/β-catenin Glu176 of human Wnt5a, Met988 of human LRP6, Gly176 (6) Zeng, X. et al. (2005) Nature 438, 873–7. pathway requires a coreceptor from proteins belonging to of human Dvl2, Leu53 of human Naked1, and Ala217 of (7) Davidson, G. et al. (2005) Nature 438, 867–72. the low-density lipoprotein receptor (LDLR) family, such human Naked2. The monoclonal antibody for Axin1 is (8) Luo, W. and Lin, S.C. Neurosignals 13, 99–113. as LRP5 and LRP6 (4). Upon stimulation with Wnt, LRP6 produced by immunizing animals with recombinant human is phosphorylated by kinases such as GSK-3 and CK1 Axin1. The phospho-specific polyclonal antibody is pro- (9) Salahshor, S. and Woodgett, J.R. (2005) J Clin Pathol 58, and subsequently recruits Axin to the membrane (5-7). In duced by immunizing animals with a synthetic phospho- 225–36. the absence of Wnt stimulation, the multidomain scaffold peptide corresponding to residues surrounding Ser1490 of (10) Boutros, M. and Mlodzik, M. (1999) Mech Dev 83, 27–37. proteins Axin1 and Axin2 negatively regulate Wnt signaling. human LRP6. The total polyclonal antibody is produced by Axin1 complexes with APC, GSK-3β, Dvl and β-catenin immunizing animals with a synthetic peptide corresponding (11) Wharton, K.A. et al. (2001) Dev Biol 234, 93–106. and promotes the GSK-3β-mediated phosphorylation and to residues surrounding the carboxy terminus of human (12) Yan, D. et al. (2001) Proc Natl Acad Sci USA 98, 3802–7. subsequent degradation of β-catenin (8-9). The Disheveled Dvl3. Polyclonal antibodies are purified by Protein A and (Dsh) proteins (Dvl1, Dvl2 and Dvl3) inhibit glycogen syn- peptide affinity chromotagraphy. thase kinase-3β, thereby promoting β-catenin stabilization (4,10). Naked inhibits the canonical Wnt/β-catenin pathway by binding to Dsh proteins and directing Dsh activity towards the planar cell polarity pathway (11-12). is a trademark of Cell Signaling Technology, Inc. is a trademark of Cell Signaling Technology, ® Applications Key: W—Western IP—Immunoprecipitation IHC—Immunohistochemistry ChIP—Chromatin Immunoprecipitation IF—Immunofluorescence F—Flow cytometry E-P—ELISA-Peptide Species Cross-Reactivity Key: H—human M—mouse R—rat Hm—hamster Mk—monkey Mi—mink C—chicken Dm—D. melanogaster X—Xenopus Z—zebra fish B—bovine ® 2014 Cell Signaling Technology, Inc. ® 2014 Cell Signaling Technology, Cell Signaling Technology Dg—dog Pg—pig Sc—S. cerevisiae All—all species expected Species enclosed in parentheses are predicted to react based on 100% homology..