PI 3-Kinase HTRF Assay

The Complete PI3-Kinase Assay Solution

	The PI3-Kinase HTRF Assay is a high-performance assay kit that provides a universal method for assaying all Class I PI3-Kinases in a homogeneous 384 well format. Nowhere else will you find a kit that offers this level of performance. And, nowhere else you will find as many Class I PI3-Kinases available. PI 3-Kinase HTRF

How does it work?

This industry-leading assay makes use of the specific, high affinity binding of the GRP1 pleckstrin homology (PH) domain to PIP₃, the product of a Class 1A or 1B PI3-Kinase acting on its physiological substrate PIP₂. During the detection phase of the assay, a complex is generated between the GST-tagged PH domain and biotinylated short chain PIP₃. The biotinylated PIP₃ and the GST-tagged PH domain recruit fluorophores (Streptavidin-Allophycocyanin and Europium-labeled anti-GST respectively) to form the fluorescence resonance energy transfer (FRET) architecture, generating a stable time-resolved FRET signal. The FRET complex can be disrupted in a competitive manner by non-biotinylated PIP₃, a product formed in the PI3-Kinase assay. And because the assay has been configured into a robust homogeneous format utilizing homogenous time-resolved fluorescence (HTRF) technology, the integrity of enzyme kinetics are preserved, ensuring the most relevant assay parameters.

Use it as a kit to screen for potential PI3-Kinase inhibitors, or optimize further for use as a stand-alone module to test compounds for their ability to inhibit the binding of PIP₃ to the PH domain. It all adds up to ultra high throughput, increased reliability and robust performance so critical in helping you research PI3-Kinases and their possible link to diseases such as cancer.

The PI3-Kinase HTRF Assay Highlights

Large Stokes shift and time-resolved elements of technology help to reduce compound interference, minimizing concern of false negatives.
Picomole sensitivity of the HTRF assay platform enables the integrity of enzyme kinetics to be retained, ensuring the most physiologically relevant assay parameters.
Compatible with all Class I PI3-Kinase isoforms, enabling relative data comparisons and eliminating the need for multiple assay types.
Reliable, proven assay—based on the technology used in the Millipore, Upstate® KinaseProfiler™ Service.
Z` factor routinely exceeds 0.8, ensuring consistent, reliable data.