By Robert Brown, Uta Böger-Brown
Prime medical and laboratory scientists describe state of the art equipment for interpreting the mechanisms of mobile resistance to anticancer cytotoxics in human tumors. The protocols comprise specified directions and broad troubleshooting suggestions that permit researchers successfully to review a large choice of drug resistance mechanisms, together with points of drug-induced telephone dying, drug uptake/efflux, drug metabolism, and DNA fix. every one strategy is designed to aid establish the correlation among molecular and biochemical facts and the medical responses of the sufferer. Cytotoxic Drug Resistance Mechanisms illuminates all of the clinically appropriate mechanisms used as markers of the organic reaction to anticancer biotherapeutics this present day.
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Extra resources for Cytotoxic Drug Resistance Mechanisms (Methods in Molecular Medicine)
14. Propidium iodide (PI) staining solution: PBS + 5 µg/mL PI (Molecular Probes, Eugene, OK) + 200 µg/mL DNase-free RNase (Sigma) make up fresh immediately before use. The RNase can be made as a stock of 20 mg/mL in H2O and stored in aliquots at –20°C. Flow cytometer: We use a Becton Dickinson FACS Vantage instrument. ). It is important, therefore, that control reactions are included in each experiment (see Note 5). It is wise to use 34 Jones and Dive Fig. 1. Incorporation of bt-dUTP by exogenous TdT into the DNA of human ovarian carcinoma cells exposed to the DNA damaging agent cisplatin.
49, 4435–4440. Cell Sensitivity Assays: TUNEL Assay 31 4 Cell Sensitivity Assays Detection of Apoptotic Cells In Vitro Using the TUNEL Assay Neil A. Jones and Caroline Dive 1. 1. Principles of Flow Cytometry Flow cytometry allows rapid multiparameter analyses of individual cells. Cells are illuminated with incident laser light of a specific wavelength. Resultant light scatter signals and fluorescence-emission signals from fluorochromes contained at the surface of the cell or within it are detected by an array of photomultiplier tubes.
1995) Further characterisation of the in situ terminal deoxynucleotidyl transferase (TdT) assay for the flow cytometric analysis of apoptosis in drug resistant and drug sensitive leukaemic cells. Cytometry 20, 245–256. 13. , and Reutelingsperger, C. (1995) A novel assay for apoptosis based upon flow cytometric detection of phosphatidylserine on the cell surface with use of FITC-labelled annexin V. Clin. Chem. 41, 91. 14. , Gregory, C. , Phipps, D. , Evans, D. , Milner, A. , and Wyllie, A. H. (1992) Analysis and discrimination of necrosis and apoptosis (programmed cell death) by multi parameter flow cytometry.
Cytotoxic Drug Resistance Mechanisms (Methods in Molecular Medicine) by Robert Brown, Uta Böger-Brown