| Assay Method Information | |
| | Kinase (PDGFRα, PDGFRβ, ABL1 and FLT3) Inhibition Test of Compounds |
| Description: | The method adopted in the test was Caliper Mobility Shift Assay, which was a detection platform based on the mobility detection technology of microfluidic chip technology. Test steps: 1.25 kinase reaction buffer (62.5 mmol/L HEPES, pH 7.5; 0.001875% Brij-35; 12.5 mmol/L MgCl2; 2.5 mM DTT) and kinase reaction stop solution (100 mmol/L HEPES, pH 7.5; 0.015% Brij-35; 0.2% Coating Reagent #3) were configured. 10 μL of 2.5 kinase solution (adding kinase in 1.25 kinase reaction buffer) was added into 5 μL of compound solution with 5 concentration (dissolved in DMSO, diluted 10 times with water), incubated at room temperature for 10 minutes, then added with 10 μL of 2.5 substrate peptide solution (adding FAM labeled peptide and ATP in 1.25 kinase reaction buffer), reacted at 28 C. for a specific time, and then added with 25 μL of kinase reaction stop solution. Collected data was tested on Caliper to yield that inhibition ratio to kinase activity=(max−conversion)/(max−min) 100. max was DMSO control without adding compound, and min was low control. |
| Affinity data for this assay | |
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