A high-throughput fluorescent polarization model for screening EPO receptor ligand / 中国药理学通报

Aim To discover lead compound whose function is similar to EPO.Methods A high-throughput Fluorescent Polarization(FP) assay was established for screening ligand of EPO receptor from small molescules and natural products.In the FP assay,EPO receptor was abstracted from FVA cell.EPO was labeled with FITC.Results The optimized concentration of FITC-EPO was 10~(-9) mol?L~(-1).Different concentrations of DMSO(VDMSO/Vtotal: 0%,0.5%,1%,2.5%,5%) didn′t affect the FP assay.The value of Z′ in the FP assay was 0.78.Conclusion The assay was robust and of high sensitivity.

HTS model for protein tyrosine kinase inhibitors / 中国药理学通报

Aim To introduce a high throughput screen model for PTK inhibitors. Methods With tyrosine containing polypeptides as substrate, in reaction buffer PTK extracts catalyzed the phosphorylation of tyrosine residues. ELISA method was used to measure the activity of protein tyrosine kinase. Resutls The IC_ 50 of genistein was 110 ?mol?L -1. From more than 7000 samples, 16 were found to possess some degree of PTK inhibiting activity. Conclusion This HTS model for screening PTK inhibitors was stable and sensitive.