Akt Activity Assay Kit, K-LISA™
from
EMD Millipore
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Product
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Akt Activity Assay Kit, K-LISA™
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Company
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EMD Millipore
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Price
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More Information
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View Company Product Page
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Detection
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Request Info
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Form
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96-well plate
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Method
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Colorimetric
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Original Item Name
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K-LISA™ Akt Activity Kit
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Range
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6-100 mU/well
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Sample Type
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Cell lysates, tissue extracts, purified enzyme
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Sensitivity
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0.05 mUnits
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Target
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Akt
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Target/Molecule Descriptor
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K-LISA™
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Time
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3 h
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Catalog Number
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CBA019-1KIT
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Quantity
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96 Tests
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Description
The K-LISA™ Akt Activity Kit is a rapid, sensitive, ELISA-based activity assay suitable for measuring the kinase activity of purified or partially purified Akt preparations, in vitro Akt inhibitor screening, and for assessing the regulation of Akt in cell signaling. The assay utilizes a biotinylated peptide substrate (GRPRTSSFAEG) that is phosphorylated on the second serine by Akt1, Akt2, Akt3, SGK (Serum Glucocorticoid Kinase), and MSK1. Biotinylated Akt substrate and sample containing Akt are incubated in the presence of ATP in wells of a streptavidin-coated 96-well plate, which allows for phosphorylation and substrate capture in a single step. The phosphorylated substrate is detected with a phosphoserine detection antibody, followed by an HRP-antibody conjugate and color development with TMB substrate. Sensitivity is increased by the addition of ELISA stop solution, and relative activity is determined by reading the absorbance at dual wavelengths 450/540 nm or 450/595 nm. Addition of inhibitor (staurosporine) serves as a negative control. Includes a streptavidin-coated 96-well plate, biotinylated Akt Substrate, phosphoserine detection antibody, antibody diluent, kinase assay buffer, ATP/MgCl2 mix, inhibitor, kinase stop solution, wash concentrate, HRP-antibody conjugate, TMB substrate, ELISA stop solution, plate sealers, and user protocol.
References
Hill, M.M. et al. 2002. Pharmacol. Ther. 93, 243.Shin, I. et al. 2002. Nat. Med. 8, 1136.Zheng, et al. 2002. J. Biol. Chem. 42, 39379.Brazil, D.P. et al. 2001. Trends Biochem. Sci. 26, 657.Medema, R.H., et al. 2000. Nature 404, 782.Datta, S.R., et al. 1999. Genes Dev. 13, 2905.Franke, F.T., et al. 1997. Cell 88, 435. Alessi, D.R., et al. 1996. EMBO J. 15, 6541.
Specifications/Features
Kit Contents:
The K-LISA™ Akt Activity Kit is a rapid, sensitive, ELISA-based activity assay suitable for measuring the kinase activity of purified or partially purified Akt preparations, in vitro Akt inhibitor screening, and for assessing the regulation of Akt in cell signaling. The assay utilizes a biotinylated peptide substrate (GRPRTSSFAEG) that is phosphorylated on the second serine by Akt1, Akt2, Akt3, SGK (Serum Glucocorticoid Kinase), and MSK1. Biotinylated Akt substrate and sample containing Akt are incubated in the presence of ATP in wells of a streptavidin-coated 96-well plate, which allows for phosphorylation and substrate capture in a single step.
AKT1
(AKT1)
PTK2B
(Focal adhesion kinase 2)