Revision 7
#8814
Store at -20C
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.
Applications:
W, IP, IHC-P, IF-IC, FC-FP, ChIP, ChIP-seq, C&R
Reactivity:
H M R Mk
Sensitivity:
Endogenous
MW (kDa):
92
Source/Isotype:
Rabbit IgG
UniProt ID:
#P35222
Entrez-Gene Id:
1499
Product Usage Information
The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.
| Application | Dilution |
|---|---|
| Western Blotting | 1:1000 |
| Immunoprecipitation | 1:100 |
| Immunohistochemistry (Paraffin) | 1:400 - 1:1600 |
| Immunofluorescence (Immunocytochemistry) | 1:800 - 1:3200 |
| Flow Cytometry (Fixed/Permeabilized) | 1:800 - 1:1600 |
| Chromatin IP | 1:100 |
| Chromatin IP-seq | 1:100 |
| CUT&RUN | 1:100 |
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 #33893.
For a carrier free (BSA and azide free) version of this product see product #33893.
Specificity/Sensitivity
Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb recognizes endogenous β-catenin protein when residues Ser33, Ser37, and Thr41 are not phosphorylated. It does not detect β-catenin protein if tri-phosphorylated at Ser33/Ser37/Thr41. This antibody may also detect β-catenin protein when singly phosphorylated at Ser33. This specificity data was derived from competition ELISA and dot blot analysis using synthetic peptides.
Species predicted to react based on 100% sequence homology
Chicken, Xenopus, Zebrafish, Bovine, Dog, Pig, Horse, Guinea Pig
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser37 of human β-catenin protein.
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).
Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb is designed to specifically recognize the stabilized form of β-catenin, i.e., protein that has not been phosphorylated by GSK-3, and thus is functionally active in cell-cell adhesion and/or the canonical Wnt signaling pathway.
Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb is designed to specifically recognize the stabilized form of β-catenin, i.e., protein that has not been phosphorylated by GSK-3, and thus is functionally active in cell-cell adhesion and/or the canonical Wnt signaling pathway.
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.
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
W: Western Blotting IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized) ChIP: Chromatin IP ChIP-seq: Chromatin IP-seq C&R: CUT&RUN
Cross-Reactivity Key
H: Human M: Mouse R: Rat Mk: Monkey
Trademarks and Patents
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
SignalStain is a registered trademark of Cell Signaling Technology, Inc.
SimpleChIP 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 7
Western blot analysis of extracts from control HeLa cells (lane 1) or β-Catenin knockout HeLa cells (lane 2) using Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). The absence of signal in the β-Catenin knockout HeLa cells confirms the specificity of the antibody for β-Catenin.
Western blot analysis of extracts from 293T cells, untreated (-) or treated (+) with LiCl (20 mM, 20 hr at 37ºC), using Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb (left) and β-Catenin Antibody #9562 (right). Equal protein loading was confirmed using β-Tubulin (9F3) Rabbit mAb #2128.
Western blot analysis of extracts from 293T cells, untreated (-) or treated (+) with MG132 (10 μM, 4 hr at 37ºC), using Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb (left), Phospho-β-Catenin (Ser33/37/Thr41) Antibody #9561 (center), or β-Catenin (6B3) Rabbit mAb #9582 (right). Note that Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb fails to detect poly-ubiquitinated β-catenin in MG132-treated cells, indicating its specificity for stabilized protein.
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 7
Immunohistochemical analysis of paraffin embedded human breast carcinoma using Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded Apc (Min/+) mouse intestinal adenoma (left) and adjacent normal intestinal epithelium (right) using Non-phospho (Active) beta-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb. Note the nuclear accumulation of active beta-Catenin in the adenoma cells.
Immunohistochemical analysis of paraffin embedded cell pellets, HeLa (left) or NCI-H28 (right), using Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) 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 7
Immunohistochemical analysis of paraffin embedded mouse colon using Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb in the presence of phospho-β-catenin (Ser33/37/Thr41) peptide (left) or non-phospho-β-catenin (Ser33/37/Thr41) peptide (right). Note the absence of staining only in the presence of the non-phospho-β-catenin (Ser33/37/Thr41) peptide, demonstrating non-phospho specificity.
Confocal immunofluorescent analysis of HCT-15 (left, center; positive) or NCI-H28 (right; negative) cells using Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb (green) in the presence of Phospho-Catenin-β (Ser33/Ser37/Thr41) peptide (left) or Non-phospho-Catenin-β (Ser33/Ser37/Thr41) peptide (center). Red = Propidium Iodide (PI)/RNase Staining Solution #4087.
Flow cytometric analysis of K562 cells, untreated (blue) or treated with MG-132 #2194 (10 µM, 4 hr; green) using Non-phospho (Active) B-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E mAb IgG XP Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor 488 Conjugate) #4412 was used as a secondary 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 7
Chromatin immunoprecipitations were performed with cross-linked chromatin from HCT116 cells and either TCF4 (C48H11) Rabbit mAb #2569 or Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. TCF4 and β-Catenin are known to associate with each other on chromatin. The figure shows binding of both TCF4 and β-Catenin across CAMK2D, a known target gene of both TCF4 and β-Catenin (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product datasheet.
Chromatin immunoprecipitations were performed with cross-linked chromatin from HCT116 cells and either TCF4 (C48H11) Rabbit mAb #2569 or Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. TCF4 and β-Catenin are known to associate with each other on chromatin. The figure shows binding of both TCF4 and β-Catenin across chromosome 4 (upper), including CAMK2D (lower), a known target gene of both TCF4 and β-Catenin (see additional figure containing ChIP-qPCR data).
Chromatin immunoprecipitations were performed with cross-linked chromatin from HCT 116 cells and either Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) 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.
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 7
CUT&RUN was performed with HCT 116 cells and Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) 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).
CUT&RUN was performed with HCT 116 cells and Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) 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 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).
CUT&RUN was performed with HCT 116 cells and either Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) 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, human c-Myc promoter primers, 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.
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.