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| Product Name | Cell Cycle and Apoptosis Analysis Kit |
| Catalog No. | RESK-0139 |
| Description | This kit uses the classic propidium iodide (PI) staining method to analyze cell cycle distribution and apoptosis. PI is a double-stranded DNA fluorescent dye that intercalates into dsDNA and emits fluorescence proportional to the DNA content. After PI staining, the DNA content of cells can be measured by flow cytometry, enabling cell cycle and apoptosis analysis based on the DNA content distribution. Assuming the fluorescence intensity of G0/G1-phase cells is 1, G2/M-phase cells containing a doubled genome show a theoretical fluorescence intensity of 2, while S-phase cells undergoing DNA replication show fluorescence intensities between 1 and 2. Because apoptotic cells undergo nuclear condensation and DNA fragmentation, part of the genomic DNA is lost during staining, so apoptotic cells are weakly stained by PI (fluorescence intensity < 1), producing the characteristic sub-G1 peak (apoptotic cell peak) on the flow cytometry histogram. Apoptosis can also be monitored by changes in light scatter. During apoptosis, cytoplasmic and chromatin condensation, nuclear fragmentation and apoptotic body formation alter light scatter properties: in early apoptosis forward scatter decreases markedly as chromatin condenses and cell density increases; in late apoptosis both forward and side scatter decrease markedly as apoptotic bodies form. The kit is generally applied to cell cycle and apoptosis analysis of adherent or suspension cells. For tissue samples, the tissue must first be digested into a single-cell suspension before analysis. |
| Intended Use | For the analysis of cell cycle distribution (G0/G1, S, G2/M) and apoptosis (sub-G1 population) in cultured adherent or suspension cells by flow cytometry. For research use only. |
| Principle / Technology | Propidium iodide (PI) intercalates into double-stranded DNA and fluoresces in proportion to DNA content. DNA content analysis distinguishes G0/G1 (fluorescence = 1), S (1-2) and G2/M (2) phase cells; apoptotic cells, owing to nuclear condensation and DNA fragmentation, lose DNA during staining and appear as a sub-G1 peak. Apoptosis-associated changes in forward/side light scatter provide an additional readout. |
| Detection Method | Flow cytometry (PI fluorescence for DNA content; forward/side scatter for apoptotic morphology) |
| Sample Type | Adherent or suspension cultured cells; tissue samples must be digested into single-cell suspensions before analysis |
| Performance Range / Specifications | Convenient and accurate |
| Specificity | PI specifically stains double-stranded DNA after RNA removal by RNase A; fluorescence intensity is proportional to dsDNA content |
| Reaction Conditions / Protocol | 1. Harvest cells and resuspend thoroughly in cold PBS to obtain a single-cell suspension. 2. Fix cells by slowly adding pre-cooled absolute ethanol to a final concentration of 70-75% (do not resuspend cells directly in 70-75% ethanol, which causes poor fixation or cell aggregation). 3. Wash, treat with RNase A, and stain with PI in Staining Solution according to the manual. 4. Analyze DNA content by flow cytometry and quantify G0/G1, S and G2/M populations; the sub-G1 peak indicates apoptotic cells. |
| Components / Formulation | Propidium Iodide (PI), 1 mg/mL RNase A, 1 mg/mL Staining Solution (salt-based buffer formulation; PBS can be used as a temporary substitute) The kit buffer does not contain ATP. |
| Storage Conditions | Store at -20°C protected from light. For repeated use within a short period, the kit may be stored at 4°C protected from light. |
| Shelf Life | 2 years when stored at -20°C protected from light; 2 months when stored at 4°C protected from light |
| Package Specifications | 50 T 100 T |
| Product Form | Liquid kit (ready-to-use staining reagents) |
| Concentration | PI: 1 mg/mL; RNase A: 1 mg/mL |
| Shipping Conditions | Shipped on wet ice |
| Expiration Date / Stability | Stable for 2 years at -20°C protected from light; stable for 2 months at 4°C protected from light for short-term repeated use |
| Regulatory / Compliance | For research use only |
| Compatibility | Compatible with standard flow cytometers equipped for PI detection |
| Recommended Buffer System | Kit Staining Solution; PBS or DPBS may be used as a substitute |
| Application Notes / Precautions | Do not fix cells by directly adding 70-75% ethanol; resuspend cells in PBS first, then add absolute ethanol to 70-75% to avoid aggregation and fixation failure. Cell loss during washing can be minimized by using conical-bottom tubes and a swing-out rotor, aspirating (not decanting) the supernatant, and leaving a thin liquid film above the pellet. A single DNA histogram peak or a missing G2/M peak may result from poor cell condition, excessive plating density (contact inhibition), non-proliferating primary cells, or improper gating. If cell cycle phase percentages do not sum to 100%, check gating to exclude doublets/clumps or adjust the fitting model. A centrifugation speed of 1000 x g is a recommendation; reduce speed or use the laboratory's routine conditions if cell death is observed. For apoptosis-suppressing or cycle-arresting drug treatments, an increased G2/M fraction with a decreased S fraction is typically observed; compare the ratios of each phase. |
For research use only, not for clinical use.
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