Akt1, PKBalpha Immunoprecipitation Kinase Assay Kit, BioAssay(TM)

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\nphosphorylation 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\npeptide can be used to assay the activity of immunoprecipitated Akt/PKB.\n \nCapacity: 25 assays \n \nComponents: \n5X ADBI\nAnti-Akt/PKB PH Domain\nAkt/SGK Speci?c Peptide\nPKA Inhibitor Peptide\nMg/ATP Cocktai\np81 Paper\n\nOther components required but not included as part of kit are:\nEnzyme preparation or stimulated cell extract-containing active Akt/PKB\n\nProtein G Agarose\n\nBuffer 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\n\nBuffer B: 50mM Tris-HCl, pH 7.5, 0.03% (w/v) Brij-35, 0.1mM EGTA and 0.1% (v/v) 2-mercaptoethanol\n\nacetone\nvortex mixer\nplexiglass shielding\nshaking incubator\ntimer\nvariable volume (5-200ul) pipet + tips\nphosphoric acid\nscintillation vials\nscintillation fluid\nscintillation counter\n[g-32P]ATP- ~3000 Ci/mmol\n\nStorage and Stability:\nStable for 1 year at -20 degrees C from date of shipment.

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SPECIFICATIONS

Catalog Number

A1125-15

Size

1Kit

Applications

IP

References

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.