PI is an intercalating red fluorescent agent that binds between the bases of DNA. Upon binding to DNA, the fluorescence of Propidium Iodide (PI) is enhanced 20 to 30 fold. PI binding shifts the maximum fluorescence excitation by about 35 nm towards red while also shifting the maximum emission by about 15 nm towards blue. One molecule of PI stoichiometrically binds every four to five base pairs of DNA.
Propidium Iodide Stain
Propidium Iodide (PI) is an intercalating fluorescent agent that binds between the bases of DNA. Propidium Iodide is membrane impermeant, which prevents DNA binding in viable cells, allowing identification of dead cells in a population.
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1. Add PI to the cell sample media at 0.5% v/v. For example, add 1.5 µL PI to 300 µL of cells.
2. Incubate 5-10 minutes at room temperature.
3. Visualize with a fluorescence microscope with excitation at 488 nm and emission at 635 nm.
4. Alternatively, cells can be analyzed with a flow cytometer using a blue laser at 488 nm or a green laser at 525 nm. Read fluorescent output in FL-2 or FL-3.
5. When bound to DNA, the maximum absorption is 535 nm and the maximum emission is 617.
• Target: necrotic and apoptotic cells
• Supplied at: 250 µg/mL
• Excitation/Emission: 535 nm / 617 nm
• pH: 4.5 – 6.0
• Method of Analysis: Flow cytometer, Fluorescence Microscope
• Storage: 2-8°C, protected from light
• Shipping: Ships overnight (domestic), International Priority Shipping