Wnt / Hedgehog / Notch

β-Catenin (L54E2) Mouse mAb (Alexa Fluor® 488 Conjugate) #2849

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Applications Key:   IF-IC = Immunofluorescence (Immunocytochemistry)  F = Flow Cytometry
Reactivity Key: H = Human  M = Mouse  Mk = Monkey  Pg = Pig  R = Rat  
Species cross-reactivity is determined by Western blot.

Protocols

2849

Flow, Immunofluorescence,

Specificity / Sensitivity

β-Catenin (L54E2) Mouse mAb (Alexa Fluor® 488 Conjugate) detects endogenous levels of total β-catenin protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the carboxy terminus of human β-catenin. The antibody was conjugated to Alexa Fluor® 488 under optimal conditions with an F/P ratio of 2-5.

Description

This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 488 fluorescent dye and tested in-house for direct flow cytometric analysis of human cells. The unconjugated antibody #2677 reacts with human, mouse, rat and monkey total β-catenin protein. CST expects that β-Catenin (L54E2) Mouse mAb (Alexa Fluor® 488 Conjugate) will also recognize β-catenin in these species.

IF-IC

Confocal immunofluorescent analysis of HeLa cells using β-Catenin (L54E2) Mouse mAb (Alexa Fluor® 488 Conjugate (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Flow Cytometry

Flow cytometric analysis of NCI-H28 cells (blue) and HeLa cells (green) using β-Catenin (L54E2) Mouse mAb (Alexa Fluor® 488 Conjugate).

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. Morin, P.J. et al. (1997) Science275, 1787 - 1790.
2. Cadigan, K.M. and Nusse, R. et al. (1997) Genes Dev.11, 3286 - 3305.
3. Wodarz, A. and Nusse, R. et al. (1998) Annu. Rev. Cell. Dev. Biol.14, 59 - 88.
4. Polakis, P. et al. (1999) Curr. Opin. Genet. Dev.9, 15 - 21.
5. Amit, S. et al. (2002) Genes Dev.16, 1066 - 1076.
6. Lin, C. et al. (2002) Cell108, 837 - 847.
7. Yanagawa, S. et al. (2002) EMBO J.21, 1733 - 1742.
8. Yost, C. et al. (1996) Genes Dev.10, 1443 - 1454.

Application References

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Companion Products

• 2560 LRP6 (C5C7) Rabbit mAb
• 2230 LEF1 (C12A5) Rabbit mAb
• 2203 TCF1 (C63D9) Rabbit mAb
• 2286 LEF1 (C18A7) Rabbit mAb

This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.