Propidium iodide solution

DNA quantification Human - MDA-MB-231

DNA quantification Human - MDA-MB-231
Propidium iodide solution from Sigma-Aldrich

Protocol tips

Protocol tips
Place aluminum foil over
the sample to protect it from light and incubate at room temperature for 5-10 minutes.

Optimize incubation time to
minimize signal bleeding through to other channels.

Publication protocol

Cell cycle analysis
MDA-MB-231 cells were seeded in duplicate into 6-well plates at 15 × 10 cells and incubated for 24 h prior to drug treatment for 24 h, or 48 h. Cells were then harvested, washed twice with PBS, centrifuged at 200 g for 5 min at 4 °C, re-suspended in 1 mL of cold PBS, fixed in 4 mL of cold absolute ethanol and then stored at −20 °C until staining and analysis. Fixed cells were then treated for 1 h with 200 μM DNase-free RNase A, stained with 1 mM propidium iodide (PI) and incubated for 10 min in the dark. Fluorescence of PI, a measure of DNA content in a cell population, was performed using flow cytometry (FACSCanto II, Becton Dickinson). A total of 10,000 gated events were acquired to assess the proportions of cells in different stages of the cell cycle. Analysis of cell cycle distribution was performed using FlowJo Software.

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4 years ago

Author: Dorota Ostrowska Poland

About nuclease-free water in DNA quantification

Hi everyone! I have a question about DNA quantification. Do I need to use nuclease-free water for DNA quantification and are there any repercussions if I don’t use any?

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Manufacturer protocol

Download the product protocol from Sigma-Aldrich for Propidium iodide solution below.

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