Revision 2
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.
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UniProt ID:

#P49841

Entrez-Gene Id:

2932

Product Includes Quantity Reactivity MW(kDa) Isotype
Phospho-GSK-3β (Ser9) (D85E12) XP® Rabbit mAb 5558 100 µl H M R Hm 46 Rabbit IgG
GSK-3β (D5C5Z) XP® Rabbit mAb 12456 100 µl H M R Mk 46 Rabbit IgG

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

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

Glycogen synthase kinase-3 (GSK-3) was initially identified as an enzyme that regulates glycogen synthesis in response to insulin (1). GSK-3 is a ubiquitously expressed serine/threonine protein kinase that phosphorylates and inactivates glycogen synthase. GSK-3 is a critical downstream element of the PI3K/Akt cell survival pathway whose activity can be inhibited by Akt-mediated phosphorylation at Ser21 of GSK-3α and Ser9 of GSK-3β (2,3). GSK-3 has been implicated in the regulation of cell fate in Dictyostelium and is a component of the Wnt signaling pathway required for Drosophila, Xenopus, and mammalian development (4). GSK-3 has been shown to regulate cyclin D1 proteolysis and subcellular localization (5).

  1. Welsh, G.I. et al. (1996) Trends Cell Biol 6, 274-9.
  2. Srivastava, A.K. and Pandey, S.K. (1998) Mol Cell Biochem 182, 135-41.
  3. Cross, D.A. et al. (1995) Nature 378, 785-9.
  4. Nusse, R. (1997) Cell 89, 321-3.
  5. Diehl, J.A. et al. (1998) Genes Dev 12, 3499-511.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    PhosphoPlus is a registered trademark of Cell Signaling Technology, Inc.
    U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
    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 2
    #8213

    PhosphoPlus® GSK-3β (Ser9) Antibody Duet

    PhosphoPlus® GSK-3β (Ser9) Antibody Duet: Image 1 Expand Image
    Simple Western™ analysis of lysates (0.1 mg/mL) from PDGF-treated 3T3 cells using Phospho-GSK-3β (Ser9) (D85E12) XP® Rabbit mAb #5558. The virtual lane view (left) shows the target band (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (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.
    PhosphoPlus® GSK-3β (Ser9) Antibody Duet: Image 2 Expand Image
    Western blot analysis of extracts from wild-type, GSK-3α (-/-), and GSK3β (-/-) mouse embryonic fibroblasts (MEFs) using GSK-3β (D5C5Z) XP® Rabbit mAb (upper) and GSK-3α/β (D75D3) XP® Rabbit mAb #5676 (lower). (MEF wild type, GSK-3α (-/-), and GSK-3β (-/-) cells were kindly provided by Dr. Jim Woodgett, University of Toronto, Canada).
    PhosphoPlus® GSK-3β (Ser9) Antibody Duet: Image 3 Expand Image
    Western blot analysis of extracts from GSK-3α (-/-) (lanes 1,2) and GSK-3β (-/-) (lanes 3,4) mouse embryonic fibroblast (MEF) cells, λ phosphatase or PDGF-treated, using Phospho-GSK-3β (Ser9) (D85E12) XP® Rabbit mAb (upper) and α/β-Tubulin Antibody #2148 (lower). (MEF wild type, GSK-3α (-/-) and GSK-3β (-/-) cells were kindly provided by Dr. Jim Woodgett, University of Toronto, Canada).
    PhosphoPlus® GSK-3β (Ser9) Antibody Duet: Image 4 Expand Image
    Immunoprecipitation of GSK-3β from PC-12 cell extracts, using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or GSK-3β (D5C5Z) XP® Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using GSK-3β (D5C5Z) XP® Rabbit mAb.
    PhosphoPlus® GSK-3β (Ser9) Antibody Duet: Image 5 Expand Image
    Western blot analysis of extracts from PC-3 cells, untreated or LY294002/wortmannin-treated, using Phospho-GSK-3β (Ser9) (D85E12) XP® Rabbit mAb (upper) or GSK-3β (27C10) Rabbit mAb #9315 (lower).
    PhosphoPlus® GSK-3β (Ser9) Antibody Duet: Image 6 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma using GSK-3β (D5C5Z) XP® Rabbit mAb.
    PhosphoPlus® GSK-3β (Ser9) Antibody Duet: Image 7 Expand Image
    Confocal immunofluorescent analysis of wild type mouse embryonic fibroblasts (MEFs) (top row), GSK-3β (-/-) MEFs (middle row) , or PC-3 cells (bottom row), untreated (left), LY294002- and Wortmannin-treated (#9901 and #9951 respectively; center) or lambda phosphatase-treated (right), using Phospho-GSK-3β (Ser9) (D85E12) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). (MEF wild type and GSK-3β (-/-) cells were kindly provided by Dr. Jim Woodgett, University of Toronto, Canada).
    PhosphoPlus® GSK-3β (Ser9) Antibody Duet: Image 8 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse pancreas using GSK-3β (D5C5Z) XP® Rabbit mAb.
    PhosphoPlus® GSK-3β (Ser9) Antibody Duet: Image 9 Expand Image
    Flow cytometric analysis of NIH/3T3 cells, untreated (blue) or PDGF-treated (green), using Phospho-GSK-3β (Ser9) (D85E12) XP® Rabbit mAb.
    PhosphoPlus® GSK-3β (Ser9) Antibody Duet: Image 10 Expand Image
    Immunohistochemical analysis of paraffin-embedded MEF cell pellets, wild type (left), GSK-3α (-/-) (middle) and GSK-3β (-/-) (right) using GSK-3β (D5C5Z) XP® Rabbit mAb. (MEF wild type, GSK-3β (-/-), and GSK-3α (-/-) cells were kindly provided by Dr. Jim Woodgett, University of Toronto, Canada).
    PhosphoPlus® GSK-3β (Ser9) Antibody Duet: Image 11 Expand Image
    Confocal immunofluorescent analysis of wild-type mouse embryonic fibroblasts (MEFs) (left), GSK-3α (-/-) MEFs (center)and GSK-3β (-/-) MEFs (right) using GSK-3β (D5C5Z) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). (MEF wild type, GSK-3α (-/-), and GSK-3β (-/-) cells were kindly provided by Dr. Jim Woodgett, University of Toronto, Canada).
    PhosphoPlus® GSK-3β (Ser9) Antibody Duet: Image 12 Expand Image
    Flow cytometric analysis of GSK-3β (-/-) MEFs (blue, negative) and wild type mouse embryonic fibroblasts (MEFs) (green, positive) using GSK-3β (D5C5Z) XP® Rabbit mAb or a 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. (MEF wild type and GSK-3β (-/-) cells were kindly provided by Dr. Jim Woodgett, University of Toronto, Canada).