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9278
GSK-3 Fusion Protein
Experimental Controls

GSK-3 Fusion Protein #9278

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Supporting Data

MW (kDa) 30

Application Key:

  • W-Western
  • IP-Immunoprecipitation
  • IHC-Immunohistochemistry
  • ChIP-Chromatin Immunoprecipitation
  • IF-Immunofluorescence
  • F-Flow Cytometry
  • E-P-ELISA-Peptide

Species Cross-Reactivity Key:

  • H-Human
  • M-Mouse
  • R-Rat
  • Hm-Hamster
  • Mk-Monkey
  • 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
  • All-All Species Expected

Product Description

GSK-3 Fusion Protein serves as a useful substrate for assaying Akt kinase activity.

Molecular Formula:

Molecular Weight: 30 kDa

Product Usage Information

GSK-3 fusion protein at a concentration of 1ug/40uL reaction is phosphorylated by an upstream kinase in an in vitro kinase assay with 1X kinase buffer #9802 and 200uM ATP #9804. After a 30 minute assay at 30C, phosphorylation can be detected using a phospho-GSK3 antibody.

Quality Control

Protein kinase substrates are examined for purity by SDS-PAGE analysis and quality control tested by Western blot using our compatible antibodies. Each substrate is also tested for its ability to be phosphorylated by its upstream kinase. This is confirmed by Western blot with the appropriate CST phospho-specific antibody.

Source / Purification

GSK-3 α/β crosstide, corresponding to residues surrounding GSK-3 α/β (Ser21/9) (CGPKGPGRRGRRRTSSFAEG), was fused to the N-terminus of paramyosin. The fusion protein was prepared by incubating the crosstide with bacterial-expressed paramyosin, using the IMPACT-CN System (New England Biolabs #E6900). Purified by the IMPACT-CN System (NEB #E6900).

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.

Pathways & Proteins

Explore pathways + proteins related to this product.

For Research Use Only. Not For Use In Diagnostic Procedures.

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

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