Recombinant human Akt3 / PKB gamma, active protein kinase 20 µg

Background: Protein kinase B or Akt (PKB/Akt) is a serine/threonine kinase, which in mammals comprises three highly homologous members known as PKB alpha (Akt1), PKB beta (Akt2) and PKB gamma (Akt3). PKB/Akt is a growth-factor-regulated protein kinase which contains a pleckstrin homology (PH) domain. Binding of phosphoinositide 3-OH kinase products to the pleckstrin homology domain results in translocation of PKB/Akt to the plasma membrane where it is activated by phosphorylation by upstream kinases including the phosphoinoside-dependent kinase 1 (PDK1). Key roles for this enzyme can be found in cellular processes such as glucose metabolism, cell proliferation, apoptosis, transcription and cell migration. Recombinant protein kinase PKB gamma expressed in Sf9 cells.

Expression system: Sf9 cells
Purification: One-step affinity purification using Ni-NTA agarose
Purity: > 95% by SDS PAGE
Theoretical MW: 60.5 kDa
Storage buffer: 50 mM Tris-HCl, 100 mM NaCl, 1 mM DTT, 50 % glycerol, pH 8.5.
Storage temperature: - 80°C (avoid repeated freeze-thaw cycles !)
Protein concentration: 1.0 mg/ml (Bradford with BSA as standard protein)
Specific activity: 467,100 U*/mg (1 Unit is defined as 1 picomole phosphate transferred to synthetic peptide (RPRAATF) per min at 30 °C)

Entrez Gene ID: 10000 
UniProtKB:  Q9Y243

Ordering information: shipped on dry ice

Product specific literature references:

Barthel A, Nakatani K, Dandekar AA, Roth RA (1998) "Protein kinase C modulates the insulin-stimulated increase in Akt1 and Akt3 activity in 3T3-L1 adipocytes" Biochem. Biophys. Res. Commun. 243(2):509-13

Masure S, Haefner B, Wesselink JJ, Hoefnagel E, Mortier E, Verhasselt P, Tuytelaars A, Gordon R, Richardson A (1999) "Molecular cloning, expression and characterization of the human serine/threonine kinase Akt-3" Eur. J. Biochem. 265(1):353-60

Brodbeck D, Cron P, Hemmings BA (1999) "A human protein kinase Bgamma with regulatory phosphorylation sites in the activation loop and in the C-terminal hydrophobic domain" J. Biol. Chem. 274(14):9133-6

Nakatani K, Sakaue H, Thompson DA, Weigel RJ, Roth RA (1999) "Identification of a human Akt3 (protein kinase B gamma) which contains the regulatory serine phosphorylation site" Biochem. Biophys. Res. Commun. 257(3):906-10

Okano J, Gaslightwala I, Birnbaum MJ, Rustgi AK, Nakagawa H (2000) "Akt/protein kinase B isoforms are differentially regulated by epidermal growth factor stimulation" J. Biol. Chem. 275(40):30934-42

Brodbeck D, Hill MM, Hemmings BA (2001) "Two splice variants of protein kinase B gamma have different regulatory capacity depending on the presence or absence of the regulatory phosphorylation site serine 472 in the carboxyl-terminal hydrophobic domain" J. Biol. Chem. 276(31):29550-8