Analysis of Cell Cycle by Flow Cytometry
Described are four widely used procedures to analyze the cell cycle by flow cytometry. The first two are based on univariate analysis of cellular DNA content following cell staining with either propidium iodide (PI) or 4′,6′-diamidino-2-phenylindole (DAPI) and deconvolution of the cellular DNA content frequency histograms. This approach reveals distribution of cells in three major phases of the cycle (G1 vs S vs G2 /M) and makes it possible to detect apoptotic cells with fractional DNA content. The third approach is based on the bivariate analysis of DNA content and proliferation-associated proteins. The expression of cyclin D, cyclin E, cyclin A, or cyclin B1 vs DNA content is presented as an example. This approach allows one to distinguish, for example, G0 from G1 cells, identify mitotic cells, or relate expression of other intracellular proteins to the cell cycle position. The fourth procedure relies on the detection of 5′-bromo-2′-deoxyuridine (BrdU) incorporation to label the DNA-replicating cells.
- T-Cell Receptor Clonotype Mapping Using Denaturing Gradient Gel Electrophoresis: Analysis of Clonal T-Cell Responses in Melanoma
- Analysis of the PML/RAR- Fusion Gene in Acute Promyelocytic Leukemia by Reverse-Transcription Polymerase Chain Reaction: Technic
- Electrophoretic Mobility Shift Analysis of the DNA Binding of Tumor Suppressor Gene Products
- DNA Methylation and Histone Modifications in Breast Cancer
- Practical Methods for Tissue Microarray Construction
- The NF-B Transcription Factor Pathway as a Therapeutic Target in Cancer: Methods for Detection of NF-B Activity
- Recombinant Single-Chain and Disulfide-Stabilized Fv Immunotoxins for Cancer Therapy
- Production of Ligand-Specific Mutants Using a Yeast Two-Hybrid Mating Assay
- Chromogenic In Situ Hybridization in Tumor Pathology
- 原位雜交的難題