Technical Data
A1125-15
Akt1, PKBalpha Immunoprecipitation Kinase Assay Kit, BioAssay™
1Kit
Kits and Assays Storage: 4°CShipping: Blue Ice
Used in the Immunoprecipitation and assessment of Akt 1 kinase activity from lysate. The sequence of this peptide (RPRAATF) is related to the sequence surrounding the
phosphorylation site of Glycogen Synthase Kinase-3 (GSK3). This peptide, which has one potential phosphorylation site, is a relatively specific substrate for Akt/PKB, because it is not phosphorylated by other protein kinases such as Rsk-2 and p70 S6 Kinase. The
peptide can be used to assay the activity of immunoprecipitated Akt/PKB.

Capacity: 25 assays

Components:
5X ADBI
Anti-Akt/PKB PH Domain
Akt/SGK Speci c Peptide
PKA Inhibitor Peptide
Mg/ATP Cocktai
p81 Paper

Other components required but not included as part of kit are:
Enzyme preparation or stimulated cell extract-containing active Akt/PKB

Protein G Agarose

Buffer A: 50mM Tris, pH 7.5, 1mM EDTA, 1mM EGTA, 0.5mM NaVO4, 0.1% (v/v) 2-mercaptoethanol, 1% Triton X-100, 50mM sodium fluoride, 5mM sodium pyrophosphate, 10mM sodium -glycerol phosphate, 0.1mM PMSF, 1ug/ml of aprotinin, pepstatin, leupeptin, and 1uM Microcystin

Buffer B: 50mM Tris-HCl, pH 7.5, 0.03% (w/v) Brij-35, 0.1mM EGTA and 0.1% (v/v) 2-mercaptoethanol

acetone
vortex mixer
plexiglass shielding
shaking incubator
timer
variable volume (5-200ul) pipet + tips
phosphoric acid
scintillation vials
scintillation fluid
scintillation counter
[g-32P]ATP- ~3000 Ci/mmol

Storage and Stability:
Stable for 1 year at -20°C from date of shipment.
Important Note: This product is intended for research use only, not for use in human, therapeutic or diagnostic applications.
Standaert et al., JBC 274: 25308, 1999 Alessi, D.R., et al., FEBS Lett. 399: 333-338, 1996. James, S.R., et al., Biochem. J. 315: 709-713, 1996. Application References: Whitlock, B., et al., J. Cell Biol. 11: 1305-1320, 2000. Gardai, B., et al., J. Biol. Chem. 277: 5236-5246, 2002.

Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.