Revision 1

#9369Store at -20C

1 Kit

(5 x 20 microliters)

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.
Product Includes Product # Quantity Mol. Wt Isotype/Source
GSK-3α (D80E6) Rabbit mAb 4337 20 µl 51 kDa Rabbit IgG
Phospho-GSK-3α (Ser21) (36E9) Rabbit mAb 9316 20 µl 51 kDa Rabbit 
Phospho-GSK-3β (Ser9) (D85E12) XP® Rabbit mAb 5558 20 µl 46 kDa Rabbit IgG
Phospho-GSK-3α/β (Ser21/9) (D17D2) Rabbit mAb 8566 20 µl 46 GSK-3beta, 51 GSK-3alpha kDa Rabbit IgG
GSK-3β (D5C5Z) XP® Rabbit mAb 12456 20 µl 46 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

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

Description

The GSK-3 Antibody Sampler Kit contains primary and secondary antibodies to perform two Western blots with each antibody.

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.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    Limited Uses

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    Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

    Revision 1
    #9369

    GSK-3 Antibody Sampler Kit

    GSK-3 Antibody Sampler Kit: 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.
    GSK-3 Antibody Sampler Kit: 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).
    GSK-3 Antibody Sampler Kit: Image 3 Expand Image
    Western blot analysis of extracts from various cell types using GSK-3α (D80E6) Rabbit mAb.
    GSK-3 Antibody Sampler Kit: Image 4 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).
    GSK-3 Antibody Sampler Kit: Image 5 Expand Image
    After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
    GSK-3 Antibody Sampler Kit: Image 6 Expand Image
    Western blot analysis of extracts from COS-7 cells, λ phosphatase- or PDGF-treated (100 μg/ml, 15 min), and PC-3 cells, untreated or λ phosphatase-treated, using Phospho-GSK-3α/β (Ser21/9) (D17D2) Rabbit mAb (upper) and GSK-3α/β (D75D3) XP® Rabbit mAb #5676 (lower).
    GSK-3 Antibody Sampler Kit: Image 7 Expand Image
    Western blot analysis of extracts from COS-7 cells, λ-phosphatase or PDGF-treated, using Phospho-GSK-3α (Ser21) (36E9) Rabbit mAb (upper) or GSK-3α Antibody #9338 (lower).
    GSK-3 Antibody Sampler Kit: Image 8 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.
    GSK-3 Antibody Sampler Kit: Image 9 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).
    GSK-3 Antibody Sampler Kit: Image 10 Expand Image
    Western blot analysis of extracts from GSK-3α (-/-) and GSK-3β (-/-) mouse embryonic fibroblasts (MEF), untreated or insulin-treated (100 ng/ml, 20 min) , using Phospho-GSK-3α/β (Ser21/9) (D17D2) Rabbit mAb (upper) and GSK-3α/β (D75D3) XP® Rabbit mAb #5676 (lower). (MEF GSK-3α (-/-) and GSK-3β (-/-) cells were kindly provided by Dr. Jim Woodgett, University of Toronto, Canada).
    GSK-3 Antibody Sampler Kit: Image 11 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma , untreated (left) or lambda phosphatase treated (right), using Phospho-GSK-3alpha (Ser 21) (36E9) Rabbit mAb.
    GSK-3 Antibody Sampler Kit: Image 12 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma using GSK-3β (D5C5Z) XP® Rabbit mAb.
    GSK-3 Antibody Sampler Kit: Image 13 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).
    GSK-3 Antibody Sampler Kit: Image 14 Expand Image
    Immunohistochemical analysis of paraffin-embedded LNCaP cells, untreated (left) or LY294002-treated (right), using Phospho-GSK-3alpha (Ser 21) (36E9) Rabbit mAb.
    GSK-3 Antibody Sampler Kit: Image 15 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse pancreas using GSK-3β (D5C5Z) XP® Rabbit mAb.
    GSK-3 Antibody Sampler Kit: Image 16 Expand Image
    Flow cytometric analysis of NIH/3T3 cells, untreated (blue) or PDGF-treated (green), using Phospho-GSK-3β (Ser9) (D85E12) XP® Rabbit mAb.
    GSK-3 Antibody Sampler Kit: Image 17 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-GSK-3α (Ser21) (36E9) Rabbit mAb in the presence of control peptide (left) or Phospho-GSK-3α (Ser21) (36E9) Blocking Peptide #1027 (right).
    GSK-3 Antibody Sampler Kit: Image 18 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).
    GSK-3 Antibody Sampler Kit: Image 19 Expand Image
    Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Phospho-GSK-3alpha (Ser 21) (36E9) Rabbit mAb.
    GSK-3 Antibody Sampler Kit: Image 20 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).
    GSK-3 Antibody Sampler Kit: Image 21 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).