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1) Product Images from "Lapatinib-Binding Protein Kinases in the African Trypanosome: Identification of Cellular Targets for Kinase-Directed Chemical Scaffolds"
Article Title: Lapatinib-Binding Protein Kinases in the African Trypanosome: Identification of Cellular Targets for Kinase-Directed Chemical Scaffolds
Journal: PLoS ONE
Figure Legend Snippet: Drug elution of T. brucei protein kinases using an ATP-affinity resin. Proteins in total cell lysate from T. brucei were bound on ATP resin. Sepharose 4B resin was used as control. Unbound proteins (Flow through) were recovered and resin was washed sequentially with buffer A and buffer A containing 1 M KCl (Panel A). Bound proteins were eluted with lapatinib (100 µM), or AEE788 (100 µM) or canertinib (100 µM) (Panel B, C and D, respectively). Proteins were visualized by silver staining. Matrix label, A is ATP-sepharose, and S is Sepharose 4B.
Techniques Used: Flow Cytometry, Silver Staining
Figure Legend Snippet: Lapatinib, AEE788 and canertinib kill bloodstream T. brucei . Bloodstream form T. brucei (initial cell density of 2×10 3 cells/ml) were cultured in 24-well plates for 48 h with either DMSO (control) or different concentrations of drug. Cells were counted with a hemocytometer and the graphs plotted. Data are mean ± standard deviation obtained from two independent experiments performed in duplicate. ( A ). Effect of lapatinib on growth of T. brucei . ( B ). Comparison of growth inhibitory effect of lapatinib on T. brucei to HeLa cells. HeLa and T. brucei cells were cultured for 48 h with variable concentrations of lapatinib. Relative cell density represents the live cells expressed as percentages of the control ( i.e. , DMSO-treated) experiment. ( C ) Trypanosome kill assay. Time-course of trypanocidal effect of lapatinib. Trypanosomes (10 6 cells/ml) were treated with DMSO (solvent) or lapatinib (10 µM) for 8 h: viable cells were counted every hour, and plotted for each time point.
Techniques Used: Cell Culture, Standard Deviation
Figure Legend Snippet: Chemical structure of lapatinib, AEE788, and canertinib.
Figure Legend Snippet: Best models of TbLBPK•drug complexes are consistent with affinity chromatography elution data. ( A ) Predicted ICM binding scores of lapatinib (red), AEE788 (green) and canertinib (magenta) in the binding pockets of the protein kinases. The dotted line represents a hypothetical cutoff for kinase elution by drug after an NAD + wash of the affinity column. ( B ) Predicted binding poses for lapatinib (red), AEE788 (green) and canertinib (magenta) to their highest affinity protein kinases; TbLBPK1, TbLBPK2, and TbCBPK1, respectively. Kinase hinge region is shown in ribbon, ligand atom placement surface is represented by a wire mesh and colored according to its binding properties.
Techniques Used: Affinity Chromatography, Binding Assay, Affinity Column