Technology Listings


Assay to Measure Kinase Phosphatase Activity

Application

Research kit capable of selectively measuring kinase/phosphatase activity within a heterogeneous sample.

Key Benefits
  • Reaction conditions can be tailored to a particular enzyme, eliminating background noise.
  • Accurate readings can be obtained using a multitude of sample types.
  • High throughput capabilities.
Technical Summary

Emory University researchers have devised methods to detect the activity of a specific kinase/phosphatase within a heterogenous sample, eliminating inefficiency and radioactivity issues that exist with current methods. This novel assay measures the binding of a TAMRA-labeled substrate to fluorescein-labeled beads via Fluorescent Resonance Energy Transfer (FRET). Our inventors have demonstrated that FRET-based phosphatase/kinase measurements are not hampered by the presence of additional phosphate modifying enzymes; simple, well-characterized modifications to the reaction conditions are sufficient to preferentially detect activity of a chosen enzyme. This is expected to significantly reduce data collection times, as the activity of many phosphate-modifying enzymes can be quickly measured simply by changing reaction buffers. In readings taken from a multitude of samples, including whole blood, cells in culture, and isolated organs, intra-assay variability is less than 10% - an advancement over existing radiolabelled detection methods.

One disadvantage to most kinase/phosphatase assays is that they suffer from high signal to noise ratios that prevent accurate measurements of enzyme activity; ideally, phosphorylated proteins are separated from non-phosphorylated proteins before use. Because FRET-based technology eliminates the need for time-consuming purification steps and permits measurements to be taken in a single well of a multi-well plate, this kit is ideal for multiple types of high throughput screens. Accelerated assay speed, high throughput capabilities, and selective detection of specific kinase/phosphatase activity present substantial advantages to existing assays used in research settings.

Developmental Stage

This assay has been used extensively in cell culture and animal models.

Publication: Roberts B, et al. Cell Calcium. 2008; 43: 515-19.

Patent Information
App Type Country Serial No. Patent No. File Date Issued Date Expire Date
Continuation United States 13/706,750 8,748,119 7/31/2008 6/10/2014 7/31/2028
Tech ID: 07075
Published: 2/24/2009
Category
Research Tools

Contact
Hyeon (Sean) Kim
Licensing Associate
Emory University
hkim70@emory.edu

Inventor(s)
Jennifer Gooch
Brian Roberts
Jan Pohl

Keywords
Assay
Immunology/Inflammation