Product name

Propidium Iodide Flow Cytometry Kit

Detection method

Fluorescent

Sample type

Adherent cells, Suspension cells

Assay type

Cell-based

Product overview

ab139418 is designed for quantitative DNA content analysis in tissue culture cells using the nucleic acid stain propidium iodide followed by flow cytometry analysis. Propidium iodide staining of DNA is the classic means of cell cycle analysis. The staining procedure takes less than 1 hour of total processing time and cells fixed in ethanol are stable for at least several weeks at 4ºC. The contents of this kit are sufficient for 200 assays.

Propidium iodide is a fluorescent molecule that binds nucleic acid with little or no sequence preference. Because Propidium iodide binds RNA as well as DNA, RNaseA (ribonuclease A) is included in this kit to digest cellular RNA and thus decrease background RNA staining from the experiment. Since Propidium iodide is membrane impermeant, ethanol is used to both fix and permeabilize cells. This kit is compatible with cells of any species that can be prepared as a single cell suspension. A flow cytometer is required for quantitative analysis.

The excitation maximum for propium idoide is 493 nm, and the emission maximum is 636 nm.

Notes

Cell cycle analysis by quantitation of DNA content was one of the earliest applications of flow cytometry. The DNA of mammalian, yeast, plant or bacterial cells can be stained by a variety of DNA binding dyes. The premise with these dyes is that they are stoichiometric i.e. they bind in proportion to the amount of DNA present in the cell. In this way cells that are in S phase will have more DNA than cells in G1. They will take up proportionally more dye and will fluoresce more brightly until they have doubled their DNA content. The cells in G2 will be approximately twice as bright as cells in G1.