Cell Signaling Technology

Product Pathways - Wnt / Hedgehog / Notch

Phospho-β-Catenin (Ser33/37/Thr41) Blocking Peptide #1120

Description

This peptide is used to block Phospho-beta-Catenin (Ser33/37/Thr41) Antibody # 9561 reactivity.

Quality Control

The quality of the peptide was evaluated by reversed-phase HPLC and by mass spectrometry. The peptide blocks Phospho-beta-Catenin (Ser33/37/Thr41) Antibody #9561 signal completely in Western blotting and immunohistochemistry.

Western Blotting

Western Blotting

Western blot analysis of whole cell lysates from NIH/3T3 cells treated with calyculin A, using Phospho-beta-Catenin (Ser33/37/Thr41) Antibody #9561 alone (left) and the same antibody preincubated with Phospho-beta-Catenin (Ser33/37/Thr41) Blocking Peptide (right).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon, using Phospho-beta-Catenin (Ser33/37/Thr41) Antibody #9561 in the presence of control peptide (left) or Phospho-beta-Catenin (Ser33/37/Thr41) 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 twice the volume of peptide as volume of antibody used in 100 μl total volume. Incubate for a minimum of 30 minutes prior to adding the entire volume to the slide. Recommended antibody dilutions can be found on the product data sheet.

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).

  1. Cadigan, K.M. and Nusse, R. (1997) Genes Dev. 11, 3286-3305.
  2. Wodarz, A. and Nusse, R. (1998) Annu. Rev. Cell. Dev. Biol. 14, 59-88.
  3. Polakis, P. (1999) Curr. Opin. Genet. Dev. 9, 15-21.
  4. Amit, S. et al. (2002) Genes Dev. 16, 1066-1076.
  5. Lin, C. et al. (2002) Cell 108, 837-847.
  6. Yanagawa, S. et al. (2002) EMBO J. 21, 1733-1742.
  7. Yost, C. et al. (1996) Genes Dev. 10, 1443-1454.
  8. Morin, P.J. (1997) Science 275, 1787-1790.

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