The control system of the cell cycle is closely linked to cell proliferation. Cell division in which one cell produces two daughter cells occurs as part of the cell cycle. The cell cycle can be analyzed by measuring the cells stained for DNA using flow cytometry, followed by analysis of the proportion of stained nuclei in cell cycle phase. The cell cycle is classified into two major phases: interphase and the mitotic (M) phase. Interphase is composed of G1 phase (preparation for DNA synthesis which occurs prior to cell division), S phase (DNA synthesis and replication), and G2 phase (a single cell with duplicated chromosomes). M phase involves mitosis and cytokinesis. Cell cycle progression is regulated by cyclins and cyclin-dependent kinases. To determine the action of anticancer drugs and others, it is important to analyze the mechanism of cell-cycle checkpoint.
Propidium iodide (PI) is generally used for cell cycle analysis using flow cytometry. Compared to this analysis method, the Cell Cycle Assay Solution Deep Red has good membrane permeability and uses the dye with high specificity for DNA, which makes it possible to analyze the cell cycle just by adding the reagent to a cell suspension. Additionally, a 633 nm laser is available in the Cell Cycle Assay Solution Deep Red, allowing you to concurrently use a highly versatile 488 nm laser.
Clearly Identify Cell Cycle Stages
Live CHO cells stained by the Cell Cycle Assay Solution Blue and Deep Red were measured by flow cytometry. Similar experiments were performed using the existing reagent for cell cycle analysis and PI staining a widely used staining technique. As a result, results obtained by the Cell Cycle Assay Solution were equivalent to PI staining results (shown below). Compared to four different products, our product obtained a sharp histogram peak in live cells.
Doxorubicin (DOX) acts to inhibit cell proliferation during G2/M phases of the cell cycle and induces cellular senescence. After adding DOX to A549 cells, higher histogram peaks for the G2/M phase (Cell Cycle Assay Solution Blue and Deep Red), induced cellular senescence (Cellular Senescence Detection Kit – SPiDER-βGal), and differences in mitochondrial membrane potential (JC-1 MitoMP Detection Kit) were observed compared to before.