usage
sufficient for 96 tests
packaging
pkg of 1 96-well plate(s)
manufacturer/tradename
Calbiochem®
storage condition
OK to freeze
assay range
sensitivity: 0.05 mU/well, standard curve range: 6-100 mU/well
input
sample type cell lysate
sample type purified enzyme(s)
sample type tissue extract(s)
detection method
colorimetric
storage temp.
−20°C
General description
This product has been discontinued.
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.
Biochem/physiol Actions
The K-LISA Akt Activity Kit is 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.
Features and Benefits
Assay Time: 3 h
Preparation Note
Prepare all reagents immediately prior to use. The following table provides reagent preparation instructions to obtain the Volume of Working Solutions (WS) required for one 8-well strip:
Table 1: Preparation of Reagants
Upon arrival store the Streptavidin-Coated 96-Well Plate at 4°C and the remaining components of the kit at -20°C.
Other Notes
Biotinylated Akt substrate, phosphoserine detector antibody, streptavidin coated 96-well plate strips, HRP-conjugated secondary antobody, serine/threonine kinase assay buffer, ATP/MgCl₂, inhibitor, wash buffer, TMB substrate, stop solution, EDTA solution, plate sealers, and a user protocol. Purified Akt is available separately (Cat. No. 124006).
Due to the nature of the Hazardous Materials in this shipment, additional shipping charges may be applied to your order. Certain sizes may be exempt from the additional hazardous materials shipping charges. Please contact your local sales office for more information regarding these charges.
Liau, S.S., et al. 2006. J. Gastrointest. Surg.10, 1254.
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. Nature404, 782.
Datta, S.R., et al. 1999. Genes Dev.13, 2905.
Franke, F.T., et al. 1997. Cell88, 435.
Alessi, D.R., et al. 1996. EMBO J.15, 6541.
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. Nature404, 782.
Datta, S.R., et al. 1999. Genes Dev.13, 2905.
Franke, F.T., et al. 1997. Cell88, 435.
Alessi, D.R., et al. 1996. EMBO J.15, 6541.
One unit is defined to the amount of enzyme required to transfer 1 nmol phosphate per min to substrate at 30° C.
Streptavidin-Coated 96-Well Plate, Akt Substrate, Biotinylated, Phosphoserine Detection Antibody, Antibody Diluent, Kinase Assay Buffer, 5X ATP/MgCl₂ Mix, 100X Inhibitor, Kinase Stop Solution, ELISA 20X Plate Wash Concentrate, HRP-Antibody Conjugate, TMB Substrate, ELISA Stop Solution, Plate Sealers, and a user protocol. Sold under license of U.S. patent 6,441,140. Manufactured by Cell Signaling.
Legal Information
*Sold under license of U.S. patent 6,441,140Manufactured by Cell Signaling
CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
Toxicity: Multiple Toxicity Values, refer to MSDS (O)
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