| Assay Method Information | |
| | PDGFRβ LanthaScreen Assay |
| Description: | Stock Solutions:Assay buffer stock contains 50 mM HEPES pH7.5, 10 mM MgCl2, 0.01% Brij-35, 1 mM EGTA.Tb-labeled inactive PDGFRβ. 3.6 μM in 50 mM HEPES, pH 7.4, 150 mM NaCl, 0.005% Tween-20 and 10% glycerol. Store at −80° C. in aliquots.Tracer 222, 50 μM in DMSO, store at −20° C.Freshly Prepared Solutions:Assay buffer. Add DTT to 2 mM and ovalbumin to 0.1 mg/mL to Assay buffer stock.Kinase-Tracer solution. Make a working solution of 0.2 nM Tb-labeled inactive PDGFRβ and 40 nM Tracer 222 in Assay buffer. Keep on ice until use.Assay Procedure:Step 1. Dispensing inhibitors: Using Echo, dispense 40 nL/well (or less) compound serial dilutions in DMSO onto the assay plate.Step 2. Dispensing Kinase-Tracer solution: Add 4 μL/well Kinase-Tracer solution. Seal the plate with optically transparent plate seal. Centrifuge at 1000 rpm for 1 min.Final concentrations of components in the assay:[Tb-PDGFRβ]=0.2 nM;[Tracer 222]=40 nM;[DMSO]≤1%.Step 3. Detection: Read TR-FRET signals after 18 hours incubation at room temperature. |
| Affinity data for this assay | |
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