What is the concentration of propidium iodide?
It is recommended to use Propidium Iodide Solution at a final concentration of 1 μg/mL. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
How long is propidium iodide stable?
Propidium Iodide Solution is supplied at ambient temperature. Upon arrival store at 4°C. The product is stable for 1 year when stored as instructed.
How does propidium iodide staining work?
Propidium iodide (PI) is a cell-impermeant DNA binding dye that can be used to stain cells and nucleic acids. PI binds to DNA by intercalating between the bases with a stoichiometry of one dye per 4-5 base pairs of DNA. As a membrane impermeant dye, PI is generally excluded from viable cells.
Where does propidium iodide stain localize in the cell?
Where does the propidium iodine stain in the cell? Nucleus of the cell.
Can propidium iodide be fixed?
Propidium Iodide is a charged substance; i.e. the positively charged propidium ion does not enter living cells. If the aim is to stain all cells, then they must be fixed in ethanol before staining.
Why does propidium iodide stain dead cells?
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. PI is an intercalating red fluorescent agent that binds between the bases of DNA.
How do you store propidium iodide?
Contents & storage Contains 1 bottle of propidium iodide(1.0 mg/mL solution in water). Store in refrigerator (2–8°C) and protect from light.
Can I use PE and PI together?
Propidium iodide (PI) is a membrane impermeant dye that is generally excluded from viable cells. Because of these spectral characteristics, PI can be used in combination with other fluorochromes excited at 488 nm such as fluorescein isothiocyanate (FITC) and phycoerythrin (PE).
What does propidium iodide measure?
Propidium iodide is used as a DNA stain in flow cytometry to evaluate cell viability or DNA content in cell cycle analysis, or in microscopy to visualize the nucleus and other DNA-containing organelles.
How do you dilute propidium iodide?
To prepare, add 1 mg PI (Propidium Iodide) to 1 ml distilled water. Store stock solution at 4 ºC (or aliquot and store at -20 ºC), protecting from light. When handled properly, solutions are stable for many years.
How do you take propidium iodide?
Add 100 μl of propidium iodide (directly from the bottle) to the 1 ml cell suspension (106 cells). Store cells at +2 to +8°C protected from light until flow-cytometric analysis. Do not store overnight. For life science research only.
Does PI stain live or dead cells?
PI can only cross compromised bacterial membranes and is therefore considered to be an indicator of membrane integrity. It stains DNA and RNA inside of dead cells or the ones with reversibly damaged membranes.
How is propidium iodide used to determine bacterial viability?
Propidium iodide (PI) is widely used for bacterial viability staining, especially since Boulos et al. (1999) published the method 1. PI can only cross compromised bacterial membranes and is therefore considered to be an indicator of membrane integrity. It stains DNA and RNA inside of dead cells or the ones with reversibly damaged membranes.
What is the excitation maximum of propidium iodide?
Once the dye is bound, its fluorescence is enhanced 20- to 30-fold, the fluorescence excitation maximum is shifted ~30–40 nm to the red and the fluorescence emission maximum is shifted ~15 nm to the blue, resulting in an excitation maximum at 535 nm and fluorescence emission maximum at 617 nm.
Is it safe to harvest propidium iodide cells?
Note: Propidium iodide is a suspected carcinogen and should be handled with care. The dye must be disposed of safely and in accordance with applicable local regulations. Harvest cells and aliquot up to 1 x 10 6 cells/100 μL into FACS tubes.
How does propidium iodide bind to double stranded DNA?
Propidium iodide (PI) is a membrane impermeant dye that is generally excluded from viable cells. It binds to double stranded DNA by intercalating between base pairs. PI is excited at 488 nm and, with a relatively large Stokes shift, emits at a maximum wavelength of 617 nm.