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TKXtra DataFile: Homogenous tyrosine kinase detection using High Efficiency Fluorescence Polarization

A Sensitive, Robust, Homogenous,and Rapid Assay
Uses High Efficiency Fluorescence Polarization (HEFP) detection method
Assays a broad range of tyrosine kinases
Requires no separation or wash steps
Available in a kit and in bulk packaging

Introduction
Screening for tyrosine kinase activity can be accelerated using High Efficiency Fluorescence Polarization (HEFP) assays. Molecular Devices TKXtra HEFP Assay Reagents provides a sensitive and rapid assay that is applicable for detecting the activity of numerous tyrosine kinases. For convenience and economy, the reagents can be purchased as a kit for assay evaluation and in bulk for high-throughput screening.


Intended Use
This product is designed to determine an increase or decrease in activity of tyrosine kinase in solution.


Principle of the Assay
The TKXtra system measures the activity of tyrosine kinases via a competitive immunoassay for the product of enzyme activityphosphotyrosine (pY)-containing peptides. The TKXtra procedure is a two-step process: first a kinase reaction is run, then a HEFP immunoassay is performed on the reaction products to quantify the amount of phosphopeptide formed.1


Kit Contents
The kit contains anti-phosphotyrosine antibody, fluorescein-labeled tracer, phosphotyrosine calibrator, and assay buffer. When used as suggested, each kit provides sufficient reagents for 1600 40-L assays.

For assay volumes between 440 L, HE microplates are recommended for optimal results.


Assay Conditions
Samples containing phosphopeptides produced by the tyrosine kinase assay are mixed with fluorescein-labeled tracer and anti-phosphotyrosine antibody. The samples are incubated at ambient te
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