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1、Graphene OxidePeptide Nanocomplex as a Versatile Fluorescence Probe of Protein Kinase Activity Based on PhosphorylationProtection against Carboxypeptidase Digestion报告人:蔡向东报告人:蔡向东nTraditional methods of assessing kinase acitivities nGraphene&Graphene Oxide2.EXPERIMENTAL SECTION &RESULTS AND DISCUSSIO
2、NnGraphite oxide,staurosporine, forskolin,CKII,IBMX,PKA,CPY,H-89,BSA(NIUXUEQING),Tris, glycerol, DTT, EDTA, and Triton X-100,FITC-pep, FITC-Ppep, FITC-kemptidenSynergy Mx multimode microplate readernexcitation wavelength of 480 nm, fluorescence emission was scanned from 500 to 700 nmFluorescence Que
3、nching of FITC Molecules and FITC-peptide by Graphene OxideAFM topographic images (top) and height profiles (bottom) of GO (A) before and (B) after incubation with the 10 M peptide substrate of CKII.Hydrolyzation of Peptide by CPY. Fluorescence responses at 520 nm emission of 10 M FITC-labeled pepti
4、de (S-pep) digested by different concentrations of CPY for 30 min and then mixed with GO.Detection of Protein Kinase Activity and Inhibition.MCF-7 Cell Culture and Lysate PreparationnMCF-7breast cancer cells+amphotericin B+penicillin+Fsk and IBMX(DMSO)nimproved Bradford protein assay dye reagent kit+BSA3Q