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.

使用に関する制限

法的な権限を与えられたCSTの担当者が署名した書面によって別途明示的に合意された場合を除き、 CST、その関連会社または代理店が提供する製品には以下の条件が適用されます。お客様が定める条件でここに定められた条件に含まれるものを超えるもの、 または、ここに定められた条件と異なるものは、法的な権限を与えられたCSTの担当者が別途書面にて受諾した場合を除き、拒絶され、 いかなる効力も効果も有しません。

研究専用 (For Research Use Only) またはこれに類似する表示がされた製品は、 いかなる目的についても FDA または外国もしくは国内のその他の規制機関により承認、認可または許可を受けていません。 お客様は製品を診断もしくは治療目的で使用してはならず、また、製品に表示された内容に違反する方法で使用してはなりません。 CST が販売または使用許諾する製品は、エンドユーザーであるお客様に対し、使途を研究および開発のみに限定して提供されるものです。 診断、予防もしくは治療目的で製品を使用することまたは製品を再販売 (単独であるか他の製品等の一部であるかを問いません) もしくはその他の商業的利用の目的で購入することについては、CST から別途許諾を得る必要があります。 お客様は以下の事項を遵守しなければなりません。(a) CST の製品 (単独であるか他の資材と一緒であるかを問いません) を販売、使用許諾、貸与、寄付もしくはその他の態様で第三者に譲渡したり使用させたりしてはなりません。また、商用の製品を製造するために CST の製品を使用してはなりません。(b) 複製、改変、リバースエンジニアリング、逆コンパイル、 分解または他の方法により製品の構造または技術を解明しようとしてはなりません。また、 CST の製品またはサービスと競合する製品またはサービスを開発する目的で CST の製品を使用してはなりません。(c) CST の製品の商標、商号、ロゴ、特許または著作権に関する通知または表示を除去したり改変したりしてはなりません。(d) CST の製品をCST 製品販売条件(CST’s Product Terms of Sale) および該当する書面のみに従って使用しなければなりません。(e) CST の製品に関連してお客様が使用する第三者の製品またはサービスに関する使用許諾条件、 サービス提供条件またはこれに類する合意事項を遵守しなければなりません。

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
Western blot analysis of extracts from various cell lines using Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Immunoprecipitation Image 1: Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP® Rabbit mAb Expand Image
Immunoprecipitation of non-phospho β-catenin (Ser45) protein from COLO 205 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP® Rabbit mAb. Western blot analysis was performed using Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP® Rabbit mAb.
Immunohistochemistry Image 1: Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded Apc (Min/+) mouse intestinal adenoma using Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP® Rabbit mAb.
Immunohistochemistry Image 2: Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP® Rabbit mAb.
Immunohistochemistry Image 3: Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded HeLa (left) or NCI-H28 (right) cell pellets using Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP® Rabbit mAb.
Immunohistochemistry Image 4: Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma using Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP® Rabbit mAb in the presence of phospho-β-Catenin (Ser45) peptide (left) or non-phospho-β-Catenin (Ser45) peptide (right).
Immunofluorescence Image 1: Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP® Rabbit mAb Expand Image
Confocal immunofluorescent analysis of mouse colon using Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP® Rabbit mAb (green). Actin filaments were labeled with DyLight 554 phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Immunofluorescence Image 1: Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP® Rabbit mAb Expand Image
Confocal immunofluorescent analysis of SW480 (left and lower right; positive) or NCI-H28 (upper right; negative) cells using Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP® Rabbit mAb (green) in the presence of phospho-β-Catenin (Ser45) peptide (lower left) or non-phospho β-Catenin peptide (lower right). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Flow Cytometry Image 1: Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP® Rabbit mAb Expand Image
Flow cytometric analysis of NCI-H28 cells (blue, negative) and SW480 cells (green, positive) using Non-phospho (Active) β-Catenin (Ser45) (D2U8Y) XP® 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.