Product Pathways - Wnt / Hedgehog / Notch
Phospho-β-Catenin (Thr41/Ser45) Blocking Peptide #1130
Description
This peptide is used to block Phospho-beta-Catenin (Thr41/Ser45) Antibody #9565 reactivity.
Quality Control
The quality of the peptide was evaluated by reversed-phase HPLC and by mass spectrometry. The peptide has been tested to block Phospho-Beta-Catenin (Thr41/Ser45) Antibody #9565 signal completely in Western blotting and immunohistochemistry.
Western Blotting
Western blot analysis of whole cell lysates from NIH/3T3 cells treated with calyculin A, using Phospho-beta-Catenin (Thr41/Ser45) Antibody #9565 alone (left) and the same antibody preincubated with Phospho-beta-Catenin (Thr41/Ser45) Blocking Peptide (right).
IHC-P (paraffin)
Immunohistochemical staining of paraffin-embedded human breast carcinoma, using Phospho-beta-Catenin (Thr41/Ser45) #9565 preincubated with irrelevant control peptide (left) and Phospho-beta-Catenin (Thr41/Ser45) Blocking Peptide (right).
Applications
Use as a blocking reagent to evaluate the specificity of antibody reactivity in Western immunoblotting and immunohistochemistry protocols.
Directions for Use
For Western immunoblotting, add 10 µl of antibody and 10 µl of blocking peptide to 10 ml of antibody dilution buffer, and incubate at room temperature for 2 hours before allowing to react with the blot.
For immunohistochemistry, add 2 µl of antibody and 10 µl of blocking peptide to 0.088 ml of buffer, and incubate at 4?C for 2 hours before allowing to react with the slides.
Background
β-catenin is a key downstream effector in the Wnt signaling pathway (1). It is implicated in two major biological processes in vertebrates: early embryonic development (2) and tumorigenesis (3). CK1 phosphorylates β-catenin on Ser45. This phosphorylation event primes β-catenin for subsequent phosphorylation by GSK-3 (4-6). GSK-3β destabilizes β-catenin by phosphorylating it at Ser33, Ser37 and Thr41 (7). Mutations in these phosphorylation sites, which result in the stabilization of β-catenin protein levels, have been found in many tumor cell lines (8).
- Cadigan, K.M. and Nusse, R. (1997) Genes Dev. 11, 3286-3305.
- Wodarz, A. and Nusse, R. (1998) Annu. Rev. Cell. Dev. Biol. 14, 59-88.
- Polakis, P. (1999) Curr. Opin. Genet. Dev. 9, 15-21.
- Amit, S. et al. (2002) Genes Dev. 16, 1066-1076.
- Lin, C. et al. (2002) Cell 108, 837-847.
- Yanagawa, S. et al. (2002) EMBO J. 21, 1733-1742.
- Yost, C. et al. (1996) Genes Dev. 10, 1443-1454.
- Morin, P.J. (1997) Science 275, 1787-1790.
Application References
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Companion Products
- 9565 Phospho-β-Catenin (Thr41/Ser45) Antibody
