How do you dissolve propidium iodide?
Propidium iodide (PI) is supplied as a crystalline solid. A stock solution may be made by dissolving the PI in the solvent of choice. PI is soluble in organic solvents such as ethanol, DMSO, and dimethyl formamide, which should be purged with an inert gas.
Does propidium iodide detect apoptosis?
Propidium iodide (PI) is widely used in conjunction with Annexin V to determine if cells are viable, apoptotic, or necrotic through differences in plasma membrane integrity and permeability1,2.
Why does propidium iodide staining indicate apoptosis?
Propidium Iodide Propidium Iodide (PI) is a fluorescent DNA-binding dye. It freely penetrates cell membranes of dead or dying cells, but is excluded from viable cells. These properties favor the use of PI for evaluation of (apoptotic) cell death or for cell cycle analysis.
What is the concentration of propidium iodide for FACS?
Cell cycle analysis. It is recommended to use Propidium Iodide Solution at a final concentration of 1 μg/mL.
Is propidium iodide soluble in water?
Solubility : Soluble in water (1 mg/ml), DMSO, and methanol (3 mg/ml). Melting Point : 220-225° C (dec.)
How long is propidium iodide stable?
The product is stable for 1 year when stored as instructed. Propidium Iodide is a potential mutagen.
What is propidium iodide used for?
Propidium iodide (PI) is a popular red-fluorescent nuclear and chromosome counterstain. Since propidium iodide is not permeant to live cells, it is also commonly used to detect dead cells in a population. PI binds to DNA by intercalating between the bases with little or no sequence preference.
What does propidium iodide stain?
Propidium iodide is a membrane-impermeable fluorescent DNA stain. It is used to stain apoptotic cells for flow cytometry, in situ hybridization, and immunohistochemistry applications Belloc et al (1994). It can also be used to differentiate between apoptotic and necrotic cell death.
Why is propidium iodide used to identify cells that are both proliferating and have undergone necrotic death?
Since propidium iodide is not permeant to live cells, it is also commonly used to detect dead cells in a population. PI binds to DNA by intercalating between the bases with little or no sequence preference.
How long is propidium iodide good for?
Propidium iodide solution (P3566) 1 mg/mL in water 2–6°C Protect from light • • When stored as directed, solution is stable at least 6 months.
Can propidium iodide be fixed?
Adding propidium iodide or ethidium bromide to unfixed cells can be used to estimate viability of the cells, but use rather less dye and measure quickly. Only dead cells will stain by this method. I agree with Berlinda. If your aim is to measure viability (live cells = unstained), then you cannot fix them.
Does propidium iodide expire?
Storage and Shelf Life: Shelf life is 48 months. Avoid prolonged exposure of propidium iodide powder and solutions to light. Use before expiry date given on the product label.
How does propidium iodide work in flow cytometry?
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.
Can propidium iodide stain live cells?
Propidium iodide is a vital stain. a live cells exclude the dye and cannot be stained. It stains only dead cells.
What color does propidium iodide stain?
red-
Dead cells are identified by red-fluorescent propidium iodide staining.
Which feature of apoptosis can be directly detected by light microscopy?
Light and electron microscopy have identified the various morphological changes that occur during apoptosis (Hacker, 2000). During the early process of apoptosis, cell shrinkage and pyknosis are visible by light microscopy (Kerr et al., 1972).
Is propidium iodide toxic to cells?
Unlike other compounds that are cell impermeant and that fluoresce when they bind DNA (e.g. propidium iodide), the CellTox™ Green Dye is non-toxic to cells and extremely stable.
What is the role of propidium iodide in the study of apoptosis?
Studies of cellular apoptosis have been significantly impacted since the introduction of flow cytometry-based methods. Propidium iodide (PI) is widely used in conjunction with Annexin V to determine if cells are viable, apoptotic, or necrotic through differences in plasma membrane integrity and permeability 1,2.
Is propidium iodide safe for FACs?
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.
What is the use of propidium iodide in flow cytometry?
The Propidium Iodide Solution is suitable for the exclusion of dead cells from flow cytometric analysis. Propidium iodide (PI) is widely used for staining and evaluation of cell death and apoptosis or for determination of DNA content in cell cycle analysis.
What is the fluorescence of propidium iodide?
Propidium iodide (PI) is a fluorescent dye that intercalates into double-stranded nucleic acid. It is excluded from viable cells, but can penetrate cell membranes of dead or dying cells. The fluorescence emission maximum for DNA-bound PI is about 615–620 nm.