Elabscience® Propidium Iodide (PI) Staining Solution is developed to identify apoptotic and necrotic cells.
Propidium Iodide (PI) is a common DNA dye that is not permeable to cell membrane. Once binding to DNA, the flurescence of PI increases by nearly 20 fold. Due to the loss of integrity of membrane, PI can enter late apoptotic or necrotic cells to stain DNA. Cells at different apoptotic stages can be distinguished by using Propidium Iodide (PI) and Annexin V.
Iodide (PI) Staining Solution
Propidium Iodide (PI) Staining Solution
Jurkat cells were treated with 1μM Camptothecin and detected by this reagent and Annexin V-FITC.
Jurkat cells were treated with 1μM Camptothecin (Left) or not (Right) for 4 h. Annexin V-FITC single-positive cells were early apoptotic cells, Annexin V-FITC and PI double-positive cells were necrotic or late apoptotic cells, and PI single-positive cells were nude nuclear cells.
1. Induce apoptosis of suspension cells with reagents of interest. Collect cell cultures and centrifuge at 300 g for 5 min, discard the supernatant. Add appropriate PBS to wash the cells, resuspend gently and count the cells.
Note: This product is only validated in suspension cells. Good cell viability is the key to the experiment. When the adherent cells are used for apoptotic detection, treatments like digestion may increase the ratio of necrotic or apoptotic cells and cause uncontrollable effects on the experimental results. Please be aware！
2. Split the cell suspension into tubes, 1-5 × 105 cells for each. Centrifuge at 300 g for 5 min, discard the supernatant. Add appropriate PBS to wash the cells and discard the supernatant. Add 500 μL of 1 × Annexin V Binding Buffer [#Cat:E-CK-A151] to resuspend the cells.
3. Add 5 µl of Annexin V-FITC[#Cat:E-CK-A111] and 5 µl of Propidium Iodide (PI) Staining Solution to each tube.
4. Gently vortex the cells and incubate at room temperature for 15-20 min in the dark.
5. Analyze the cells immediately with proper machine settings. Otherwise, place the cells on ice in the dark and analyze within 1 h.
Store at 4°C for up to one year in dark.
1. For maximal assay performance, this reagent should be used within 12 months. Avoid freeze / thaw cycles.
2. Detect apoptosis as soon as possible after staining to avoid the increase number of apoptosis or necrosis
3. Avoid extended exposure of the samples to direct light to protect the fluorophores from quenching.
4. For your safety and health, please wear the lab coat and disposable gloves before the experiments.