Revision 6
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IHC-P, IF-IC, FC-FP

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

92

Source/Isotype:

Rabbit IgG

UniProt ID:

#P35222

Entrez-Gene Id:

1499

Product Information

Product Usage Information

This product is the carrier free version of product #8814. All data were generated using the same antibody clone in the standard formulation which contains BSA and glycerol.

This formulation is ideal for use with technologies requiring specialized or custom antibody labeling, including fluorophores, metals, lanthanides, and oligonucleotides. It is not recommended for ChIP, ChIP-seq, CUT&RUN or CUT&Tag assays. If you require a carrier free formulation for chromatin profiling, please contact us. Optimal dilutions/concentrations should be determined by the end user.

Formulation

Supplied in 1X PBS, BSA and Azide Free.

For standard formulation of this product see product #8814.

Storage

Store at -20°C. This product will freeze at -20°C so it is recommended to aliquot into single-use vials to avoid multiple freeze/thaw cycles. A slight precipitate may be present and can be dissolved by gently vortexing. This will not interfere with antibody performance.

Specificity / Sensitivity

Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb (BSA and Azide Free) 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 Reactivity:

Human, Mouse, Rat, Monkey

Species predicted to react based on 100% sequence homology

Chicken, D. melanogaster, Xenopus, Bovine, Dog

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 #8814 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.

  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.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Applications Key

WB: Western Blotting IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
SignalStain is a 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

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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

Revision 6
#33893

Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb (BSA and Azide Free)

Western Blotting Image 1: Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb (BSA and Azide Free) Expand Image
使用 Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb(上图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图)对对照 HeLa 细胞(泳道 1)或 β-连环蛋白敲除型 HeLa 细胞(泳道 2)的提取物进行蛋白质印迹分析。β-Catenin敲除型 HeLa 细胞中没有信号,这证实了抗体对 β-Catenin的特异性。数据的生成使用的是该产品的常规组分产品。
Western Blotting Image 2: Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb (BSA and Azide Free) Expand Image
使用 Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb(左)、Phospho-β-Catenin (Ser33/37/Thr41) Antibody #9561(中)或 β-Catenin (6B3) Rabbit mAb #9582(右),对未经处理 (-) 或经 MG132 处理(10 μM,在 37ºC 下处理 4 小时;+)的 293T 细胞提取物进行蛋白质印迹分析。注意,Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb 未能在 MG132 处理的细胞中检测到聚泛素化 β-链蛋白,表明对稳定蛋白有特异性。数据的生成使用的是该产品的常规组分产品。
Western Blotting Image 3: Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb (BSA and Azide Free) Expand Image
使用 Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb(左)和 β-Catenin Antibody #9562(右),对未经处理 (-) 或经 LiCl 处理(20 mM,在 37ºC 下处理 20 小时)的 293T 细胞提取物进行蛋白质印迹分析。使用 β-Tubulin (9F3) Rabbit mAb #2128 确认蛋白质负荷是否相等。数据的生成使用的是该产品的常规组分产品。
Immunohistochemistry Image 1: Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb (BSA and Azide Free) Expand Image
使用 Non-phospho (Active) beta-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb,对石蜡包埋的 Apc (Min/+) 小鼠小肠腺癌(左)和周围正常小肠上皮细胞(右)进行免疫组织化学分析。注意,活性 β-链蛋白在腺癌细胞的胞核中聚集。数据的生成使用的是该产品的常规组分产品。
Immunohistochemistry Image 2: Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb (BSA and Azide Free) Expand Image
使用 Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb 对石蜡包埋的人乳腺癌进行免疫组织化学分析。数据的生成使用的是该产品的常规组分产品。
Immunohistochemistry Image 3: Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb (BSA and Azide Free) Expand Image
在磷酸化 β-链蛋白 (Ser33/37/Thr41) 肽(左)或非磷酸化 β-链蛋白 (Ser33/37/Thr41) 肽(右)存在的情况下,使用 Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb 对石蜡包埋的小鼠结肠进行免疫组织化学分析。注意,仅在存在非磷酸化 β-链蛋白 (Ser33/37/Thr41) 肽存在的情况下才缺乏染色,表明具有非磷酸化特异性。数据的生成使用的是该产品的常规组分产品。
Immunohistochemistry Image 4: Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb (BSA and Azide Free) Expand Image
使用 Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb,对石蜡包埋的 HeLa(左)或 NCI-H28(右)细胞沉淀物进行免疫组织化学分析。数据的生成使用的是该产品的常规组分产品。
Immunofluorescence Image 1: Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb (BSA and Azide Free) Expand Image
在磷酸化链蛋白-β (Ser33/Ser37/Thr41) 肽(左)或非磷酸化链蛋白-β (Ser33/Ser37/Thr41) 肽(中)存在的情况下,使用 Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb(绿色),对 HCT-15(左、中;阳性)或 NCI-H28(右;阴性)细胞进行共聚焦免疫荧光分析。红色 = Propidium Iodide (PI)/RNase Staining Solution #4087。数据的生成使用的是该产品的常规组分产品。
Flow Cytometry Image 1: Non-phospho (Active) β-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb (BSA and Azide Free) Expand Image
使用 Non-phospho (Active) B-Catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb(实线)或浓度匹配的 Rabbit (DA1E mAb IgG XP Isotype Control #3900(虚线),对未经处理(蓝色)或已经 MG-132 #2194(10μM,4小时;绿色)处理的 K562 细胞进行流式细胞分析。使用 Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor 488 Conjugate) #4412 作为二抗。数据的生成使用的是该产品的常规组分产品。