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2951
β-Catenin Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

β-Catenin Antibody Sampler Kit #2951

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β-Catenin Antibody Sampler Kit: Image 1

Chromatin immunoprecipitations were performed with cross-linked chromatin from HCT116 cells and either β-Catenin (D10A8) XP® Rabbit mAb or Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb #8814, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across AXIN2, a known target gene of β-Catenin (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.

β-Catenin Antibody Sampler Kit: Image 2

Confocal immunofluorescent analysis of HeLa (left) and NCI-H28 (right) cells using β-Catenin (D10A8) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

β-Catenin Antibody Sampler Kit: Image 3

Western blot analysis of extracts from control HeLa cells (lane 1) or HeLa cells with an apparent in-frame truncation mutation in the gene encoding β-Catenin (lane 2) using β-Catenin (D10A8) XP® Rabbit mAb, #8480 (upper) or β-actin (D6A8) Rabbit mAb #8457 (lower). The change in β-Catenin molecular weight in the mutated HeLa cells is consistent with an in-frame deletion.

β-Catenin Antibody Sampler Kit: Image 4

Western blot analysis of extracts from 293 cells, pretreated with 20 mM LiCl for 30 minutes and then with 50 nM calyculin A, using Phospho-β-Catenin (Ser33/37/Thr41) Antibody (upper) or β-Catenin Antibody #9562 (lower).

β-Catenin Antibody Sampler Kit: Image 5

Western blot analysis of extracts from 293 cells pretreated with 20 mM LiCl for 30 minutes and then with 50 nM calyculin A, using Phospho-β-Catenin (Thr41/Ser45) Antibody (upper) or β-Catenin Antibody #9562 (lower).

β-Catenin Antibody Sampler Kit: Image 6

Western blot analysis of extracts from SK-N-MC and NTERA-2 cells, untreated or λ phosphatase-treated, using

Phospho-β-Catenin (Ser552) (D8E11) Rabbit mAb (upper) or β-Catenin (6B3) Rabbit mAb #9582 (lower).

β-Catenin Antibody Sampler Kit: Image 7

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.

β-Catenin Antibody Sampler Kit: Image 8

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).

β-Catenin Antibody Sampler Kit: Image 9

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

β-Catenin Antibody Sampler Kit: Image 10

Chromatin immunoprecipitations were performed with cross-linked chromatin from HCT 116 cells and either β-Catenin (D10A8) XP® Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human Axin2 Intron 1 Primers #8973, SimpleChIP® Human CaMK2D Intron 3 Primers #5111, human c-Myc promoter primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

β-Catenin Antibody Sampler Kit: Image 11

Western blot analysis of extracts from SW480 cells using Phospho-β-Catenin (Ser33/37/Thr41) Antibody.

β-Catenin Antibody Sampler Kit: Image 12

Western blot analysis of extracts from SW480 cells using Phospho-β-Catenin (Thr41/Ser45) Antibody.

β-Catenin Antibody Sampler Kit: Image 13

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).

β-Catenin Antibody Sampler Kit: Image 14

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).

β-Catenin Antibody Sampler Kit: Image 15

Flow cytometric analysis of NCI-H28 cells (blue) and HeLa cells (green) using β-Catenin (D10A8) XP® Rabbit mAb (solid lines) or concentration-matched Rabbit Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as secondary antibody.

β-Catenin Antibody Sampler Kit: Image 16

Confocal immunofluorescent analysis of mouse colon using β-Catenin (D10A8) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

β-Catenin Antibody Sampler Kit: Image 17

Immunohistochemical analysis of frozen mouse colon using β-Catenin (D10A8) XP® Rabbit mAb.

β-Catenin Antibody Sampler Kit: Image 18

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using β-Catenin (D10A8) XP® Rabbit mAb.

β-Catenin Antibody Sampler Kit: Image 19

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using β-Catenin (D10A8) XP® Rabbit mAb.

β-Catenin Antibody Sampler Kit: Image 20

Immunohistochemical analysis of paraffin-embedded cell pellets, HeLa (left) or NCI-H28 (right), using β-Catenin (D10A8) XP® Rabbit mAb.

β-Catenin Antibody Sampler Kit: Image 21

Immunohistochemical analysis of paraffin-embedded mouse colon using β-Catenin (D10A8) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).

β-Catenin Antibody Sampler Kit: Image 22

Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using ß-Catenin (D10A8) XP® Rabbit mAb.

β-Catenin Antibody Sampler Kit: Image 23

Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using ß-Catenin (D10A8) XP® Rabbit mAb.

β-Catenin Antibody Sampler Kit: Image 24

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using β-Catenin (D10A8) XP® Rabbit mAb.

β-Catenin Antibody Sampler Kit: Image 25

Western blot analysis of extracts from various cell lines using β-Catenin (D10A8) XP® Rabbit mAb.

To Purchase # 2951T
Product # Size Price
2951T
1 Kit  (5 x 20 µl) $ 391

Product Description

The β-Catenin Antibody Sampler Kit provides an economical means of detecting total β-catenin as well as β-catenin phosphorlylated at various residues. The kit contains enough primary and secondary antibody to perform two Western blots with each antibody.

Specificity / Sensitivity

Each antibody in this kit recognizes only its specific target and does not cross-react with other family members.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser33, Ser37 and Thr41 of human β-catenin or residues surrounding Thr41 and Ser45 of human β-catenin. Polyclonal antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro714 of human β-catenin protein, a synthetic phosphopeptide corresponding to residues surrounding Ser552 of human β-catenin protein, or 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).

Both Akt and PKA were shown to phosphorylate β-catenin at Ser552 and Ser675. Phosphorylation at Ser552 and Ser675 induces β-catenin accumulation in the nucleus and increases its transcriptional activity (9-12).

  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, 9971-9976.
  10. Fang D et al. (2007) J Biol Chem 282, 11221–9
  11. He, X.C. et al. (2007) Nat. Genet. 39, 189-198.
  12. Hino S et al. (2005) Mol Cell Biol 25, 9063–72

Pathways & Proteins

Explore pathways + proteins related to this product.

Limited Uses

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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's 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's 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.

For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.