Revision 1
#2951
Store at -20C
β-Catenin Antibody Sampler Kit
1 Kit
(5 x 20 microliters)
Orders:
877-616-CELL (2355)
Support:
877-678-TECH (8324)
3 Trask Lane | Danvers | Massachusetts | 01923 | USA
For Research Use Only. Not for Use in Diagnostic Procedures.
| Product Includes | Product # | Quantity | Mol. Wt | Isotype/Source |
|---|---|---|---|---|
| β-Catenin (D10A8) XP® Rabbit mAb | 8480 | 20 µl | 92 kDa | Rabbit IgG |
| Phospho-β-Catenin (Ser33/37/Thr41) Antibody | 9561 | 20 µl | 92 kDa | Rabbit |
| Phospho-β-Catenin (Thr41/Ser45) Antibody | 9565 | 20 µl | 92 kDa | Rabbit |
| Phospho-β-Catenin (Ser552) (D8E11) Rabbit mAb | 5651 | 20 µl | 92 kDa | Rabbit IgG |
| Phospho-β-Catenin (Ser675) (D2F1) XP® Rabbit mAb | 4176 | 20 µl | 92 kDa | Rabbit IgG |
| Anti-rabbit IgG, HRP-linked Antibody | 7074 | 100 µl | Goat |
Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.
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.
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.
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).
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).
Background References
- Cadigan, K.M. and Nusse, R. (1997) Genes Dev 11, 3286-3305.
- Wodarz, A. and Nusse, R. (1998) Annu Rev Cell Dev Biol 14, 59-88.
- Polakis, P. (1999) Curr Opin Genet Dev 9, 15-21.
- Amit, S. et al. (2002) Genes Dev 16, 1066-76.
- Liu, C. et al. (2002) Cell 108, 837-47.
- Yanagawa, S. et al. (2002) EMBO J 21, 1733-42.
- Yost, C. et al. (1996) Genes Dev 10, 1443-54.
- Morin, P.J. et al. (1997) Science 275, 1787-90.
- Taurin, S. et al. (2006) J. Biol. Chem. 281, 9971-9976.
- Fang, D. et al. (2007) J. Biol. Chem. 282, 11221-11229.
- He, X.C. et al. (2007) Nat. Genet. 39, 189-198.
- Hino, S. et al. (2005) Mol. Cell Biol. 25, 9063-9072.
Trademarks and Patents
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.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.
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Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] •
Web:
cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.
Revision 1
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.
Immunoprecipitation of Phospho-β-Catenin (Ser675) protein from SW620 + Calyculin A #9902 (50nM, 30 min) cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Phospho-β-Catenin (Ser675) (D2F1) XP® Rabbit mAb. Western blot analysis was performed using Phospho-β-Catenin (Ser675) (D2F1) XP® Rabbit mAb.
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).
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] •
Web:
cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.
Revision 1
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.
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.
CUT&RUN was performed with HCT 116 cells and β-Catenin (D10A8) XP® Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA library was prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across Axin2, a known target gene of β-Catenin (see additional figure containing CUT&RUN-qPCR data).
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] •
Web:
cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.
Revision 1
Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using ß-Catenin (D10A8) XP® Rabbit mAb performed on the Leica® BOND™ Rx.
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).
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).
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] •
Web:
cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.
Revision 1
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).
Western blot analysis of extracts from various cell lines using β-Catenin (D10A8) XP® Rabbit mAb.
CUT&RUN was performed with HCT 116 cells and β-Catenin (D10A8) XP® Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 17 (upper), including Axin2 (lower), a known target gene of β-Catenin (see additional figure containing CUT&RUN-qPCR data).
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] •
Web:
cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.
Revision 1
Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using ß-Catenin (D10A8) XP® Rabbit mAb performed on the Leica® BOND™ Rx.
Western blot analysis of extracts from SW480 cells using Phospho-β-Catenin (Ser33/37/Thr41) Antibody.
Western blot analysis of extracts from SW480 cells using Phospho-β-Catenin (Thr41/Ser45) Antibody.
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] •
Web:
cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.
Revision 1
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).
CUT&RUN was performed with HCT 116 cells and either β-Catenin (D10A8) XP® Rabbit mAb or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human Axin2 Intron 1 Primers #8973, SimpleChIP® Human CaMK2D Intron 3 Primers #5111 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.
Immunohistochemical analysis of paraffin-embedded human serous adenocarcinoma of the ovary using ß-Catenin (D10A8) XP® Rabbit mAb performed on the Leica® BOND™ Rx.
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] •
Web:
cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.
Revision 1
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).
Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using ß-Catenin (D10A8) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using β-Catenin (D10A8) XP® Rabbit mAb.
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] •
Web:
cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.
Revision 1
Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using ß-Catenin (D10A8) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using β-Catenin (D10A8) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using β-Catenin (D10A8) XP® Rabbit mAb.
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] •
Web:
cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.
Revision 1
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).
Immunohistochemical analysis of paraffin-embedded cell pellets, HeLa (left) or NCI-H28 (right), using β-Catenin (D10A8) XP® Rabbit mAb.
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).
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] •
Web:
cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.
Revision 1
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).
Flow cytometric analysis of NCI-H28 cells (green) and HeLa cells (blue) 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.
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 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across AXIN2, a known target gene of β-Catenin (see additional figure containing ChIP-qPCR data).
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] •
Web:
cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.
Revision 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 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across chromosome 17 (upper), including AXIN2 (lower), a known target gene of β-Catenin (see additional figure containing ChIP-qPCR data).
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.
Immunoprecipitation of β-Catenin from HeLa cell extracts. Lane 1 is 10% input, lane 2 is precipitated with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is β-Catenin (D10A8) XP® Rabbit mAb, #8480. Western blot was performed using β-Catenin (15B8) Mouse mAb, #37447.
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] •
Web:
cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.
Revision 1
Simple Western™ analysis of lysates (0.1 mg/mL) from HeLa cells using β-Catenin (D10A8) XP® Rabbit mAb #8480. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] •
Web:
cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.