Akt1, unactivated (v-akt Mouse Thymoma Viral Oncogene Homolog 1, C-AKT, MGC99656, PKB, PRKBA, Protein kinase B, RAC, RAC-ALPHA, RAC-alpha serine/threonine-protein kinase, RAC-PK-alpha) Recombinant

The serine/threonine kinase Akt1 has also been described as Rac-PKa or PKBa. Rac protein kinase family members feature a pleckstrin homology (PH) domain at the amino terminus and a protein-serine/threonine kinase catalytic domain at the carboxy terminus. The PH domain (amino acids 1-106) has been implicated in interactions with other proteins such as G-protein beta gamma subunits, as well as phosphoinositides. The kinase domain is located between residues 148-411. These enzymes are activated by diverse ligands such as PGDF, EGF and basic FG in NIH 3T3, Rat-1 or Swiss-3T3 cells. Akt1 is a protein of 480 amino acids and has been observed to show different migratory patterns on a western blot according to the state of phosphorylation of the protein. Phosphatase treatment has been shown to result in inactivation of the protein.\n\nN-terminus His6-tagged recombinant full length human Akt1, expressed by baculovirus in Sf21 insect.\n\nAkt1/PKBa Activation and Phosphorylation Assay: 10ng of recombinant active PDK1 was used to phosphorylate Akt1/PKBa, unactive. The activated Akt1/PKBa in turn phosphorylates the Akt specific substrate peptide, (RPRAATF). The activated Akt1/PKBa can also phosphorylate Crosstide, RPRAATF, GSKa (Ser21), GSK3b (Ser9) and Bad (Ser136). \n\nPDK1 Kinase Assay Data: PDK1 was used to phosphorylate unactive Akt1/PKBa in the presence of PIP3 and phospholipids. The activated Akt1/PKBa phosphorylates the Akt specific substrate peptide (RPRAATF).\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage, store at -20 degrees C. Aliquots are stable for at least 6 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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