Cell Signaling Technology

Product Pathways - Development

Wnt Signaling Antibody Sampler Kit #2915

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
Wnt3a (C64F2) Rabbit mAb #2721 40 µl W IP M (H) (R) 42 Rabbit IgG
Wnt5a/b (C27E8) Rabbit mAb #2530 40 µl W H M 45 Rabbit IgG
Phospho-LRP6 (Ser1490) Antibody #2568 40 µl W IP H (M) 180, 210 Rabbit
Dvl2 (30D2) Rabbit mAb #3224 40 µl W IP H M R Mk 90 to 95 Rabbit IgG
LRP6 (C47E12) Rabbit mAb #3395 40 µl W IP H M (R) 180, 210 Rabbit IgG
Dvl3 Antibody #3218 40 µl W IP H M R Hm Mk Mi B 88 to 93 Rabbit
Naked1 (C30F10) Rabbit mAb #2201 40 µl W IP H 59, 61 kDa Rabbit IgG
Naked2 (C67C4) Rabbit mAb #2073 40 µl W IP H 59, 61 Rabbit
Axin1 (C76H11) Rabbit mAb #2087 40 µl W IP IHC-P H M 110 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl Goat

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey  Mi=Mink  B=Bovine
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

Wnt3a (C64F2) Rabbit mAb detects transfected Wnt3a protein in L cells and the conditioned medium from L/Wnt3a cells. Phospho-LRP6 (Ser1490) Antibody detects endogenous levels of LRP6 protein when phosphorylated at Ser1490. The remaining antibodies in the Wnt Signaling Sampler Kit detect endogenous levels of their respective total proteins.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Naked2 (C67C4) Rabbit mAb #2073.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Axin1 (C76H11) Rabbit mAb #2087.

Western Blotting

Western Blotting

Western blot analysis of total cell lysates from HepG2 and SW620 cells using Naked1 (C30F10) Rabbit mAb #2201.


Western Blotting

Western Blotting

Western blot analysis of total cell lysates from HeLa, LN18, PANC1 cells and conditioned medium (CM) from HeLa cells, using Wnt5a/b (C27E8) Rabbit mAb #2530.

Western Blotting

Western Blotting

Western blot analysis of total cell lysates from HeLa cells untreated, treated 5 hours with conditioned medium (CM) from L cells, or treated 5 hours with conditioned medium from L cells overexpressing Wnt3a, using Phospho-LRP6 (Ser1490) Antibody #2568.

Western Blotting

Western Blotting

Western blot analysis of total cell lysates and conditioned medium (CM) from L cells overexpressing mouse Wnt3a, compared with parental L cells, using Wnt3a (C64F2) Rabbit mAb #2721.


Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Dvl3 Antibody #3218.

Western Blotting

Western Blotting

Western blot analysis of total cell lysates from various cell lines using Dvl2 (30D2) Rabbit mAb #3224.

Western Blotting

Western Blotting

Western blot analysis of extracts from Hep G2 and HeLa cells using LRP6 (C47E12) Rabbit mAb #3395.


Description

The Wnt Signaling Antibody Sampler Kit provides an economical means of detecting integral proteins within the Wnt signaling pathway. The kit contains enough primary and secondary antibody to perform four Western blots with each.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu256 of human Wnt3a, Glu176 of human Wnt5a, Met988 of human LRP6, Gly176 of human Dvl2, Leu53 of human Naked1, and Ala217 of human Naked2. The monoclonal antibody for Axin1 is produced by immunizing animals with recombinant human Axin1. The phospho-specific polyclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser1490 of human LRP6. The total polyclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding the carboxy terminus of human Dvl3. Polyclonal antibodies are purified by Protein A and peptide affinity chromotagraphy.

Background

The Wnt family includes several secreted glycoproteins that play important roles in animal development (1). Aberrant activation of Wnt signaling pathways is involved in several types of cancers (2). Wnt members bind to the Frizzled family of proteins and activate several signaling pathways (3). The canonical Wnt/β-catenin pathway requires a coreceptor from proteins belonging to the low-density lipoprotein receptor (LDLR) family, such as LRP5 and LRP6 (4). Upon stimulation with Wnt, LRP6 is phosphorylated by kinases such as GSK-3 and CK1 and subsequently recruits Axin to the membrane (5-7). In the absence of Wnt stimulation, the multidomain scaffold proteins Axin1 and Axin2 negatively regulate Wnt signaling. Axin1 complexes with APC, GSK-3β, Dvl and β-catenin and promotes the GSK-3β-mediated phosphorylation and subsequent degradation of β-catenin (8-9). The Disheveled (Dsh) proteins (Dvl1, Dvl2 and Dvl3) inhibit glycogen synthase 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).

  1. Cadigan, K.M. and Nusse, R. (1997) Genes Dev 11, 3286-305.
  2. Giles, R.H. et al. (2003) Biochim Biophys Acta 1653, 1-24.
  3. Kohn, A.D. and Moon, R.T. Cell Calcium 38, 439-46.
  4. Logan, C.Y. and Nusse, R. (2004) Annu Rev Cell Dev Biol 20, 781-810.
  5. Tamai, K. et al. (2000) Nature 407, 530-5.
  6. Zeng, X. et al. (2005) Nature 438, 873-7.
  7. Davidson, G. et al. (2005) Nature 438, 867-72.
  8. Luo, W. and Lin, S.C. Neurosignals 13, 99-113.
  9. Salahshor, S. and Woodgett, J.R. (2005) J Clin Pathol 58, 225-36.
  10. Boutros, M. and Mlodzik, M. (1999) Mech Dev 83, 27-37.
  11. Wharton, K.A. et al. (2001) Dev Biol 234, 93-106.
  12. Yan, D. et al. (2001) Proc Natl Acad Sci U S A 98, 3802-7.

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