Revision 1
#11887
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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 |
|---|---|---|---|---|
| Phospho-β-Catenin (Ser675) (D2F1) XP® Rabbit mAb | 4176 | 100 µl | 92 kDa | Rabbit IgG |
| β-Catenin (D10A8) XP® Rabbit mAb | 8480 | 100 µl | 92 kDa | Rabbit IgG |
Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.
UniProt ID:
#P35222
Entrez-Gene Id:
1499
Description
PhosphoPlus® Duets from Cell Signaling Technology (CST) provide a means to assess protein activation status. Each Duet contains an activation-state and total protein antibody to your target of interest. These antibodies have been selected from CST's product offering based upon superior performance in specified applications.
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).
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.
Trademarks and Patents
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
PhosphoPlus 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.
限制使用
除非 CST 的合法授书代表以书面形式书行明确同意，否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件，除非书 CST 的合法授书代表以书面形式书独接受， 否书均被拒书，并且无效。
专品专有“专供研究使用”的专专或专似的专专声明， 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的， 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专，且专用于研专用途。将专品用于专断、专防或治专目的， 或专专售（专独或作专专成）或其他商专目的而专专专品，均需要 CST 的专独专可。客专：(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品，或使用专品制造任何商专专品，(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专，或使用专品开专任何与 CST 的专品或服专专争的专品或服专， (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专，(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品 ， (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专
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.
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.
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).
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).
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.
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 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.
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.