Propidium Iodide Stain

Name: Propidium Iodide Stain
Brand: ImmunoChemistry Technologies
SKU: 638
Price: 23.9 USD
Availability: InStock

Propidium Iodide 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.

Figure 1. Propidium iodide excitation and emission spectra.

Figure 2. HL-60 cells were treated with a drug, then stained with FLICA® FAM-VAD-FMK poly-caspase inhibitor reagent (green) (Cat. 92) and propidium iodide (red) (PI). Scanning laser cytometer analysis. Four populations of cells were detected: (A) Unstained live cells, (B) red necrotic cells (PI), (C) green and red late apoptotic cells (FAM-FLICA® plus PI), and (D) green early apoptotic cells (FAM-FLICA®). Data courtesy of Dr. Z. Darzynkiewicz, Brander Cancer Center, NY.

Figure 3. Cell death in primary rat hippocampal neurons. (D) is a composite of FLICA® FAM-DEVD-FMK (B), PI (C), and Hoechst (A). DNA is labeled in 4 cells (A), 3 cells are caspase-positive (B), 2 are membrane-compromised (C). One cell is in early apoptosis (green but not red). Two FLICA®-positive cells are also PI-positive (red, C) (becoming membrane compromised) and are in the late stages of apoptosis rather than necrosis. Data courtesy of Dr. Z. Kahraman Akozer, University of Maryland.

Figure 1. Propidium iodide excitation and emission spectra.

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Data Sheet Safety Data Sheet

SKU: 638

Size: 1 mL

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Price:

$23.90

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Ships: 1-2 business days

Product Details

Scientific Background

Propidium iodide (PI, Catalog 638) is an intercalating red fluorescent reagent that binds between the base pairs of DNA in membrane-compromised cells and is used to identify necrotic and apoptotic cells. As PI is membrane impermeant, it cannot reach the DNA in viable cells, thus allowing the identification of cells with permeabilized membranes in a population. PI distinguishes between living and dead cells by counterstaining nucleic acids red in necrotic, dead, apoptotic and membrane-compromised cells, while the DNA in healthy cells remains unstained. Propidium iodide can be used with ICT’s FAM-FLICA® caspase inhibitor reagents (such as Catalog 92) to identify four populations of cells: living; early apoptotic; late apoptotic; and necrotic (Figures 2 and 3).

Propidium iodide is provided ready-to-use at 250 µg/mL. Just add it to the cell culture media, incubate for a few minutes, and analyze. One molecule of PI stoichiometrically binds every four to five base pairs of DNA. Unbound PI excites at 488-492 nm and exhibits an emission maximum at 635 nm. Upon binding to DNA, the fluorescence of PI is enhanced 20-30 fold. When bound to nucleic acids, the maximum absorption is 535 nm and the maximum emission is 617 nm (Figure 1). Cells may be viewed through a fluorescence microscope or analyzed with a flow cytometer. PI is for research use only. Not for use in diagnostic procedures.

Target

Necrotic and apoptotic cells

Excitation / Emission

535 nm / 617 nm

Method of Analysis

Flow cytometry, Fluorescence microscope

4.0 + 1.0

Supplied At

250 µg/mL

Usage Statement

Intended for research use or for further manufacturing into in-vitro diagnostics reagents only. Not intended for use in human or therapeutic purposes.

Country of Origin

United States

Storage

2-8°C

Shipping

Domestic: Overnight Delivery; International: Priority Shipping

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.
Visualize with a fluorescence microscope with excitation at 488 nm and emission at 635 nm.
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.
When bound to DNA, the maximum absorption is 535 nm and the maximum emission is 617.

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Propidium Iodide Stain

SKU: 638

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