DNA Content Detection Kit (Cell Cycle)
DNA Content Detection Kit (Cell Cycle)
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| Usage | 1. Induce cell apoptosis using an appropriate method. Simultaneously, establish a negative control group and collect cells. 2. Wash cells once with PBS, collect by centrifugation at 1500 rpm for 5 minutes, adjust the cell concentration to 1 × 106/mL, and prepare 1 mL of single-cell suspension. 3. After centrifugation of the prepared single-cell suspension, remove the supernatant and fix the cells with 500 μL of 70% pre-cooled ethanol for 2 hours to overnight. Store at 4°C. Wash the fixative with PBS before staining. If necessary, filter the cell suspension once through a 200-mesh cell sieve. 4. Add 100 μL of RNase A solution to the cell pellet, resuspend the cells, and incubate at 37°C in a water bath for 30 minutes. 5. Add 400 μL of PI staining solution, mix thoroughly, and incubate at 4°C in the dark for 30 minutes. 6. Analyze red fluorescence at an excitation wavelength of 488 nm. | ||||||||||||
| Description | The cell cycle refers to the entire process that a continuously dividing cell undergoes from the end of one mitotic division to the end of the next. During this process, the cell's genetic material replicates and doubles, and at the end of division, it is evenly distributed between the two daughter cells. The cell cycle can be divided into interphase and mitosis. Interphase is often divided into the resting phase (G0), the prophase of DNA synthesis (G1), the DNA synthesis phase (S), and the late phase of DNA synthesis (G2). The entire cycle can be represented as G1 →S →G2 →M. DNA cycle analysis can be used to monitor the status of each phase of the cell cycle, that is, the state of cell proliferation. This method utilizes the ability of intracellular DNA to bind to fluorescent dyes (such as propidium iodide (PI). Different DNA content in cells at different stages of the cycle, resulting in different binding of fluorescent dyes, and different fluorescence intensities detected by flow cytometry. During cell apoptosis, cytoplasm and chromatin condense, nuclear lysis occurs, and apoptotic bodies form, resulting in changes in the cells' light scattering properties. In the early stages of apoptosis, the cells' forward-angle light scattering ability decreases significantly, while their 90° angle light scattering ability increases or remains unchanged. In the late stages of apoptosis, both forward-angle and 90° angle light scattering signals decrease. Therefore, apoptotic cells can be observed by flow cytometry by measuring changes in light scattering. When cells are stained with PI, a population of cells with low DNA staining appears before the normal G0/G1 population due to reduced total DNA content. This sub-diploid peak (sub-G1) appears before the G1 peak, representing the apoptotic population. This kit can be used to monitor DNA content (cell cycle) in cultured cells (suspension and adherent). Product composition:
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| General Notes | 1. Protect propidium iodide (PI) staining solution from light during storage and use. 2. PI is toxic; wear gloves when handling and avoid contamination. 3. All fluorescent dyes are susceptible to quenching, so it is recommended that testing be completed the same day after staining. 4. For your safety and well-being, please wear a lab coat and disposable gloves during operation. |
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| Storage Temp. | -20℃, avoid light, valid for 12 months. |
