Propidium Iodide Stain

11 Citations

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.
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.

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Data Sheet Safety Data Sheet

SKU: 638

Size: 1 mL
Price:
$22.75

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.

Necrotic and apoptotic cells
535 nm / 617 nm
Flow cytometry, Fluorescence microscope
2-8°C
Domestic: Overnight Delivery; International: Priority Shipping
4.0 + 1.0
250 µg/mL
United States
  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.
  2. Visualize with a fluorescence microscope with excitation at 488 nm and emission at 635 nm.
  3. 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.
  4. When bound to DNA, the maximum absorption is 535 nm and the maximum emission is 617.

Product Specific References

PMID Publication
38098075Killinger, M., et al. 2023. Microfluidic device for enhancement and analysis of osteoblast differentiation in three-dimensional cell cultures. Journal of biological engineering, 77.
37182453Yan, C., et al. 2023. Endoplasmic reticulum stress promotes caspase-1-dependent acinar cell pyroptosis through the PERK pathway to aggravate acute pancreatitis. International immunopharmacology, 110293.
35086419Lapaquette, P., et al. 2022. Membrane protective role of autophagic machinery during infection of epithelial cells by Candida albicans. Gut microbes, 2004798.
35137097Uribe, P., et al. 2022. Autophagy is activated in human spermatozoa subjected to oxidative stress and its inhibition impairs sperm quality and promotes cell death. Human reproduction (Oxford, England), .
35462787Dai, M., et al. 2022. The Modulation of Interferon Regulatory Factor-1 via Caspase-1-Mediated Alveolar Macrophage Pyroptosis in Ventilator-Induced Lung Injury. Mediators of inflammation, 1002582.
35545676LaRock, D.L., et al. 2022. Group A Streptococcus induces GSDMA-dependent pyroptosis in keratinocytes. Nature, 527-531.
36069386Tian, J., et al. 2022. Calycosin represses AIM2 inflammasome-mediated inflammation and pyroptosis to attenuate monosodium urate-induced gouty arthritis through NF-κB and p62-Keap1 pathways. Drug development research, .
36155068Shu, L., et al. 2022. PHLDA1 promotes sevoflurane-induced pyroptosis of neuronal cells in developing rats through TRAF6-mediated activation of Rac2. Neurotoxicology, 140-151.
36271147Wu, R., et al. 2022. Mechanisms of CD40-dependent cDC1 licensing beyond costimulation. Nature immunology, .
36194675Ventura, P.M.O., et al. 2022. Concomitant deletion of Ptpn6 and Ptpn11 in T cells fails to improve anticancer responses. EMBO reports, e55399.
33527027Ge, X., et al. 2021. PARK2 attenuates house dust mite-induced inflammatory reaction, pyroptosis and barrier dysfunction in BEAS-2B cells by ubiquitinating NLRP3. American journal of translational research, 326-335.

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