Cell Cycle Staining Kit
PI staining accurately distinguishes cell-cycle phases, with assays completed in as little as 40 minutes.
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Instruction manual- Product Description
- Composition
- Precautions
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Product Overview
DNA content analysis accounts for a substantial proportion of flow cytometry. Using this approach, researchers can proceed…
Conduct toxicity assessments of anticancer drugs and analyze the prognostic outcomes and progression status of malignant tumor cells. In cell cycle analysis, DNA content is also widely employed as a key analytical parameter. By measuring cellular DNA content, one can calculate the percentage of cells in each phase of the cell cycle; additionally, methods such as DNA fragmentation assays and dual‑color DNA staining can be used to detect apoptosis. The Cell Cycle Assay Kit enables rapid cell cycle analysis with simple, straightforward操作. This product utilizes propidium iodide (PI) as a fluorescent DNA dye, allowing flow cytometry to determine the proportions of G0/G1, G2, and S phase cells within the cell cycle.
Storage conditions
Store at 2–8°C, protected from light. Transport with ice packs.
Instructions for Use
Method 1—A rapid assay that does not require cell fixation
1. Adjust the cell density to 1–5 × 10^6 cells/mL.
2. Transfer 1 mL of the cell suspension into a 1.5-mL microcentrifuge tube, centrifuge at 1,000 × g for 3–5 minutes, and discard the supernatant.
3. Add 1 mL Assay buffer and 10 μL Permeabilization solution.
4. Vortex the sample for 5–10 seconds to mix thoroughly. Incubate at room temperature in the dark for 30 minutes, then immediately proceed with flow cytometric analysis.
For observation under a fluorescence microscope, centrifuge the cells, discard the supernatant, and resuspend in fresh buffer. Add 1 drop carefully.
Place the cell suspension onto a microscope slide, cover it with a coverslip, and observe using an appropriate filter.
Method 2—Detection Method for Fixed Cells
1. Adjust the cell density to 1–5 × 10^6 cells/mL.
2. Transfer 1 mL of the cell suspension into a 1.5-mL microcentrifuge tube and centrifuge at 1,000×g for 3–5 minutes.
3. Discard the supernatant, and add 1 mL of 70% ethanol stored at −20°C to the cell pellet.
4. Vortexing disperses the cells; fix at 4°C for 2 hours—fixation for 12–24 hours may yield better results.
5. Centrifuge at 1,000×g for 3–5 minutes, then remove the ethanol.
6. Add 1 mL of PBS buffer to wash the cells, then centrifuge at 1,000×g for 3 minutes.
7. After discarding the supernatant, add 1 mL of Assay buffer.
8. Vortex the mixture to dissociate the cells, then incubate in the dark at 37°C for 30 minutes. Subsequently, store at 4°C or on ice, protected from light.
It is recommended to perform flow cytometry within 24 hours after staining, with ideal completion on the same day.
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Product Composition
Component (50 times) Assay buffer 50 mL Permeabilization solution 500 μL
Keywords
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Goonie Biotechnology (Guangzhou) Co., Ltd.
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