Safe. Non-radioactive assay.Sensitive and accurate. As low as 0.01 U/L kinase can be quantified.Homogeneous and convenient. "Mix-incubate-measure" type assay. The whole assay involves adding a single working reagent and incubation for 10 min at room temperature.Robust and amenable to HTS: Assay can tolerate up to 300 uM ATP and 10% dimethylsulfoxide (DMSO). Z factors of > 0.6 are routinely observed in 96/384-well plates. Can be readily automated on HTS liquid handling systems for tens of thousands of assays per day.
Intended Use
FL530/585nm
Detection Principle
0.01 U/L
Shelf Life
6 months
Specifity
Rapid homogenous assay for protein kinase activity and high-throughput screen for kinase inhibitors.
Reference
Liu, K et al. (2020). Different p53 genotypes regulating different phosphorylation sites and subcellular location of CDC25C associated with the formation of polyploid giant cancer cells. Journal of Experimental & Clinical Cancer Research: CR, 39(1), 83. Assay: Kinase in cell cycle-related kinases.
Hirooka, K., Kodoi, Y., Satomura, T., & Fujita, Y. (2016). Regulation of the rhaEWRBMA operon involved in l-rhamnose catabolism through two transcriptional factors, RhaR and CcpA, in Bacillus subtilis. Journal of bacteriology, 198(5), 830-845. Assay: Kinase in B. subtilis (bateria) protein.
Zhang, X., Zhang, S., Wang, J., Zi, J., Wang, J., Chen, S., & Wan, Y. (2016). Expression, Purification, and Characterization of a Sucrose Nonfermenting 1-Related Protein Kinases 2 of Arabidopsis thaliana in E. coli-Based Cell-Free System. BioMed research international 2016:9469356. Assay: Kinase in E. coli snrk2.1 protein.
Marcin, M et al (2014). Analogs of Cinnamic Acid Benzyl Amide As Nonclassical Inhibitors of Activated JAK2 Kinase. Curr Cancer Drug Targets. 14(7):638-51. Assay: Kinase in CABA analogs cells.
Mielecki, M et al (2014). Analogs of Cinnamic Acid Benzyl Amide As Nonclassical Inhibitors of Activated JAK2 Kinase. Curr Cancer Drug Targets. 14(7):638-51. Assay: Kinase in human enzymes.
Wei, J et al (2014). Soluble Expression and Purification of the Catalytic Domain of human Vascular Endothelial Growth Factor Receptor 2 in Escherichia coli. Journal of Microbiology and Biotechnology 25(8): 1227-33. Assay: Kinase in E. coli cells.
Wei, J et al (2014). Soluble Expression and Purification of the Catalytic Domain of human Vascular Endothelial Growth Factor Receptor 2 in Escherichia coli. Journal of Microbiology and Biotechnology 25(8): 1227-33. Assay: VEGFR-2-CD tyrosine kinase expressed in E. coli.
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