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, IP, IHC-P, IF-F, IF-IC, FC-FP

REACTIVITY:

H M R Mk Z Dg

SENSITIVITY:

Endogenous

MW (kDa):

92

Source/Isotype:

Rabbit IgG

UniProt ID:

#P35222

Entrez-Gene Id:

1499

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Immunohistochemistry (Paraffin) 1:500 - 1:2000
Immunofluorescence (Frozen) 1:1600 - 1:6400
Immunofluorescence (Immunocytochemistry) 1:1600 - 1:6400
Flow Cytometry (Fixed/Permeabilized) 1:800 - 1:3200

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 #24710.

Specificity / Sensitivity

Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP® Rabbit mAb recognizes endogenous levels of β-catenin protein only when Ser45 is not phosphorylated.

Species Reactivity:

Human, Mouse, Rat, Monkey, Zebrafish, Dog

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser45 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 (Ser45) (D2U8Y) XP® Rabbit mAb specifically recognizes β-catenin that is non-phosphorylated at Ser45. Non-phospho β-Catenin (Ser45) protein is useful as a readout of stabilized β-catenin protein, and therefore functionally active in cell-cell adhesion and mediating transcriptional activity via the canonical Wnt signaling pathway (9).

  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.
  9. Sakanaka, C. (2002) J Biochem 132, 697-703.

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

WB: Western Blotting IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin) IF-F: Immunofluorescence (Frozen) 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.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
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

Revision 6
#19807

Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP® Rabbit mAb

Western Blotting Image 1: Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP® Rabbit mAb Expand Image
使用 Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP®8457 Rabbit mAb(上)和 β-Actin (D6A8) Rabbit mAb #(下)对不同细胞系提取物进行蛋白质印迹分析。
Immunoprecipitation Image 1: Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP® Rabbit mAb Expand Image
对 COLO 205 细胞提取物的非磷酸化 β-链蛋白 (Ser45) 进行免疫沉淀。泳道 1 为 10% input,泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 为 Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP® Rabbit mAb。使用 Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP® Rabbit mAb 进行蛋白质印迹分析。
Immunohistochemistry Image 1: Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP® Rabbit mAb Expand Image
使用 Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP® Rabbit mAb,对石蜡包埋的 Apc (Min/+) 小鼠小肠癌进行免疫组织化学分析。
Immunohistochemistry Image 2: Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP® Rabbit mAb Expand Image
使用 Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP® Rabbit mAb,对石蜡包埋的人结肠癌进行免疫组织化学分析。
Immunohistochemistry Image 3: Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP® Rabbit mAb Expand Image
使用 Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP® 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。
Immunohistochemistry Image 4: Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP® Rabbit mAb Expand Image
在添加 phospho-β-连环蛋白 (Ser45) 肽(左图)或 non-phospho-β-连环蛋白 (Ser45) 肽(右图)的情况下,使用 Non-phospho (活性) β-连环蛋白 (Ser45) (D2U8Y) XP® 兔单克隆抗体 对石蜡包埋的人卵巢癌细胞进行免疫组织化学分析。
Immunofluorescence Image 1: Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP® Rabbit mAb Expand Image
使用 Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP® Rabbit mAb(绿色)对小鼠结肠进行共聚焦免疫荧光分析。肌动蛋白丝用 DyLight 554 phalloidin #13054(红色)标记。蓝色伪彩 = DRAQ5® #4084(DNA 荧光染料)。
Immunofluorescence Image 1: Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP® Rabbit mAb Expand Image
在 phospho-β-Catenin (Ser45) peptide(左下)或 non-phospho β-Catenin peptide(右下)存在的情况下,使用 Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP® Rabbit mAb(绿色),对 SW480(左和右下;阳性)或 NCI-H28(右上;阴性)细胞进行共聚焦免疫荧光分析。肌动蛋白微丝由 DyLight 554 Phalloidin #13054(红色)标记。蓝色伪彩 = DRAQ5® #4084(DNA 荧光染料)。
Flow Cytometry Image 1: Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP® Rabbit mAb Expand Image
使用 Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP® Rabbit mAb(实线)或浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(虚线)对 NCI-H28 细胞(蓝色,阴性)和 SW480 细胞(绿色,阳性)进行流式细胞分析。Anti-rabbit IgG (H+L)、F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 作为二抗。