Revision 4
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IF-F, IF-IC

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

92

Source/Isotype:

Rabbit IgG

UniProt ID:

#P35222

Entrez-Gene Id:

1499

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:200
Immunofluorescence (Frozen) 1:100 - 1:200
Immunofluorescence (Immunocytochemistry) 1:100 - 1:200

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier free (BSA and azide free) version of this product see product #68076.

Specificity / Sensitivity

Phospho-β-Catenin (Ser675) (D2F1) XP® Rabbit mAb detects endogenous levels of β-catenin only when phosphorylated at Ser675.

Species Reactivity:

Human, Mouse, Rat

Species predicted to react based on 100% sequence homology

Mouse, Rat, Chicken, Xenopus, Zebrafish

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser675 of human β-catenin.

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 at Ser45. This phosphorylation event primes β-catenin for subsequent phosphorylation by GSK-3β (4-6). GSK-3β destabilizes β-catenin by phosphorylating it at Ser33, Ser37, and Thr41 (7). Mutations at these sites result in the stabilization of β-catenin protein levels and have been found in many tumor cell lines (8).

PKA was shown to phosphorylate β-catenin at Ser675. Phosphorylation at Ser675 induces β-catenin accumulation in the nucleus and increases its transcriptional activity (9,10).

  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, 1066-76.
  5. Liu, C. et al. (2002) Cell 108, 837-47.
  6. Yanagawa, S. et al. (2002) EMBO J 21, 1733-42.
  7. Yost, C. et al. (1996) Genes Dev 10, 1443-54.
  8. Morin, P.J. et al. (1997) Science 275, 1787-90.
  9. Taurin, S. et al. (2006) J Biol Chem 281, 9971-6.
  10. Hino, S. et al. (2005) Mol Cell Biol 25, 9063-72.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IF-F: Immunofluorescence (Frozen) IF-IC: Immunofluorescence (Immunocytochemistry)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 4
#4176

Phospho-β-Catenin (Ser675) (D2F1) XP® Rabbit mAb

Western Blotting Image 1: Phospho-β-Catenin (Ser675) (D2F1) XP® Rabbit mAb Expand Image
Western blot analysis of extracts from control HeLa cells (lane 1) or HeLa cells with a targeted mutation in the gene encoding β-Catenin (lane 2) using Phospho-β-Catenin (Ser675) (D2F1) XP® Rabbit mAb (upper), or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). The change in β-Catenin molecular weight in the mutated HeLa cells confirms the specificity of the antibody for β-Catenin.
Western Blotting Image 2: Phospho-β-Catenin (Ser675) (D2F1) XP® Rabbit mAb Expand Image
Western blot analysis of extracts from MKN-45 and SK-N-MC cells, untreated or treated with λ phosphatase for 1 hour or forskolin (FSK) for 30 minutes, using Phospho-β-Catenin (Ser675) (D2F1) XP® Rabbit mAb (upper) or β-Catenin (6B3) Rabbit mAb #9582 (lower).
No image available
Immunofluorescence Image 1: Phospho-β-Catenin (Ser675) (D2F1) XP® Rabbit mAb Expand Image
Confocal immunofluorescent analysis of rat colon using Phospho-β-Catenin (Ser675) (D2F1) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 Phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Immunofluorescence Image 1: Phospho-β-Catenin (Ser675) (D2F1) XP® Rabbit mAb Expand Image
Confocal immunofluorescent analysis of HeLa cells, untreated (left), λ phosphatase-treated (middle), or untreated NCI-H28 cells (β-catenin null; right) using Phospho-β-Catenin (Ser675) (D2F1) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).