ProFluor Src-Family Kinase Assay
"Add-Mix-Measure" Convenience for High-Throughput Src-Family Kinase Screening
The ProFluor® Src-Family Kinase Assay measures the activity of purified Src-family tyrosine kinases (Src, Lck, Lyn, Fyn, and Hck tested) in a multiwell plate format and involves "add-mix-read" steps only—ideal for high-throughput applications.
The assay begins with a standard kinase reaction performed with a provided Src-family kinase bisamide rhodamine 110 peptide substrate. Following the kinase reaction, a termination buffer containing a protease reagent is added, which simultaneously stops the kinase reaction and removes amino acids specifically from the nonphosphorylated substrate, liberating highly fluorescent rhodamine 110. Phosphorylated substrate, however, is resistant to digestion by the protease reagent and remains nonfluorescent. Thus, fluorescence intensity measured in this assay is inversely correlated with kinase activity. A control peptide (AAF-AMC) is included to control for compounds that may inhibit the protease and reduce false-positive hits.
Minimal Test Compound InterferenceRhodamine 110 fluorescent signal produced is much higher than the fluorescent signal given off by test compounds. The assay produces excellent Z´ values (>0.7) in either 96- or 384-well plate formats and easily distinguishes known Src-family kinase inhibitors from other compounds.
ProFluor® Src-Family Kinase Assay Technical Bulletin
PDF (422 KB)
Certificate of AnalysisLookup Certificate of Analysis
-30C TO -10C
For Research Use Only. Not for Use in Diagnostic Procedures.