EnzyChrom™ Kinase Assay Kit

Protocol SDS

Application

• Rapid homogenous assay for protein kinase activity and high-throughput screen for kinase inhibitors.

Key Features

• 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 µM 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.

Method

• FL530/585nm

Samples

• All kinases

Species

• All

Procedure

• 10 min

Size

• 400 tests

Detection Limit

• 0.01 U/L

Shelf Life

• 6 months

More Details

Kinases, also known as phosphotransferases, constitute a large family of enzymes that transfer phosphate groups from the high-energy donor molecule ATP to their specific substrates. Kinases are known to regulate the majority of cellular processes. The largest group of this family is the protein kinases. So far, 518 different kinases have been identified in humans, and up to 30% of human proteins are modified by these kinases. The enormous diversity and their key role in cellular signaling make them ideal targets for drug development. BioAssay Systems’ EnzyChrom™ Kinase Assay Kit provides a simple and rapid method for assaying kinase activity and high-throughput screening for kinase inhibitors. This homogeneous microplate-based assay involves incubating the kinase with a single working reagent, in which ADP is enzymatically converted to ATP and pyruvate, which is quantified using a fluorimetric (530nm/590nm) assay method.

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For more detailed product information and questions, please feel free to email us. Or for more general information regarding our assays, please refer to Technical Support.

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.

To find more recent publications, please click here.

If you or your labs do not have the equipment or scientists necessary to run this assay, BioAssay Systems can perform the service for you.

– Fast turnaround
– Quality data
– Low cost

Please email or call 1-510-782-9988 x 2 to discuss your projects.