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2951
β-Catenin Antibody Sampler Kit

β-Catenin Antibody Sampler Kit #2951

Chromatin IP-seq 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 from 5 ng enriched ChIP DNA using NEBNext® Ultra™ II DNA Library Prep Kit for Illumina®, and sequenced on the Illumina NextSeq. 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.

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IF-IC 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).

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Western Blotting Image 3

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

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Western Blotting 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).

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Western Blotting 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).

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Western Blotting 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).

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Western Blotting Image 7

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).

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Western Blotting Image 8

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.

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Chromatin IP Image 9

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.

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IHC-P (paraffin) Image 10

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

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Western Blotting Image 11

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

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Western Blotting Image 12

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

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IF-IC Image 13

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).

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IHC-P (paraffin) Image 14

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

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IF-F Image 15

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).

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IHC-P (paraffin) Image 16

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

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IHC-P (paraffin) Image 17

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).

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IHC-P (paraffin) Image 18

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

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IHC-F (frozen) Image 19

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

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Flow Cytometry Image 20

Flow cytometric analysis of NCI-H28 (blue) or HeLa (green) cells using β-Catenin (D10A8) XP® Rabbit mAb.

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IF-F Image 21

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).

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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
β-Catenin (D10A8) XP® Rabbit mAb 8480 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
  • ChIP
H M R Mk 92 Rabbit IgG
Phospho-β-Catenin (Ser33/37/Thr41) Antibody 9561 20 µl
  • WB
H M R Mk 92 Rabbit 
Phospho-β-Catenin (Thr41/Ser45) Antibody 9565 20 µl
  • WB
H M Mk 92 Rabbit 
Phospho-β-Catenin (Ser552) (D8E11) Rabbit mAb 5651 20 µl
  • WB
  • IP
H M 92 Rabbit IgG
Phospho-β-Catenin (Ser675) (D2F1) XP® Rabbit mAb 4176 20 µl
  • WB
  • IP
  • IF
H 92 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

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.

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

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.

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, 3286-305.
  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. Fang D et al. (2007) J Biol Chem 282, 11221–9
  10. Hino S et al. (2005) Mol Cell Biol 25, 9063–72
  11. Taurin, S. et al. (2006) J. Biol. Chem. 281, 9971-9976.
  12. He, X.C. et al. (2007) Nat. Genet. 39, 189-198.
Entrez-Gene Id
1499
Swiss-Prot Acc.
P35222
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.

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