Propidium Iodide Ribonuclease
Propidium Iodide Ribonuclease - Related Videos
09:09
Annexin V and Propidium Iodide Labeling
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2023
Staining with annexin V and propidium iodide (PI) provides researchers with a way to identify different types of cell death—either necrosis or apoptosis. This technique relies on two components. The first, annexin V, is a protein that binds certain phospholipids called phosphatidylserines, which normally occur only in the inner, cytoplasm-facing leaflet of a cell’s membrane, but become “flipped” to the outer leaflet during the early stages of apoptosis. The second component is the DNA-binding...
07:34
Modified Annexin V/Propidium Iodide Apoptosis Assay For Accurate Assessment of Cell Death
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Cited by 543 •
2011
An accurate method for the assessment of cell death is described. The protocol improves upon conventional Annexin V/ propidium iodide (PI) protocols, which display up to 40% false- positive events in cell lines and primary cells from a broad range of animal models.
04:31
Differential Nuclear Staining Assay: An Assay to Determine Mitocan Cytotoxicity by Propidium Iodide and Hoechst Double Staining
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2025
This video demonstrates a nuclear staining method to detect the cytotoxic effect of mitocan on drug-sensitive cancer cells and healthy variants. The double staining with DNA-binding dyes such as propidium iodide and Hoechst dye provides a clear distinction between the dead and live cells, thus helping to assess the cytotoxicity of the drug.
08:53
Assessment of Neuronal Viability Using Fluorescein Diacetate-Propidium Iodide Double Staining in Cerebellar Granule Neuron Culture
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Cited by 29 •
2017
This protocol describes how to accurately measure neuronal viability using Fluorescein diacetate (FDA) and Propidium Iodide (PI) double staining in cultured cerebellar granule neurons, a primary neuronal culture used as an in vitro model in neuroscience and neuropharmacology research.
10:20
Rapid Production of Recombinant Human SLFN14 Ribonuclease and Stoichiometric Analysis by Mass Photometry
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2026
This protocol describes the production of recombinant human SLFN14 ribonuclease and its stoichiometric analysis by mass photometry.
FAQs
Ribonuclease removes cellular RNA before fluorescence is measured. Without this digestion step, RNA could contribute to the nucleic-acid staining signal and interfere with the relationship between fluorescence and DNA abundance. Removing that contribution helps flow cytometry produce DNA-content profiles that more clearly separate populations associated with different cell-cycle states.
The measured signal reflects the amount of propidium iodide bound to cellular DNA, so cells with different DNA contents produce distinguishable fluorescence populations. Flow-cytometric profiles can therefore associate groups of cells with G0/G1, S, or G2/M phases. The same measurement can also reveal sub-G1 DNA or populations with abnormal ploidy.
Fixation and permeabilization prepare cells for intracellular DNA staining by preserving the cellular sample and allowing the ribonuclease and propidium iodide reagents to access nucleic acids. These preparation steps precede RNA digestion and DNA labeling, creating the conditions needed for fluorescence measurements that represent cellular DNA content rather than an untreated cell suspension.
A typical workflow begins by fixing and permeabilizing the cells. Ribonuclease is then used to digest RNA, followed by propidium iodide staining of the remaining DNA. The prepared sample is analyzed by flow cytometry, which records fluorescence from individual cells and organizes the measurements into DNA-content populations for interpretation.
Sub-G1 and abnormal-ploidy populations identify DNA-content patterns that fall outside the expected major cell-cycle groups. Their presence can signal an altered cellular DNA-content profile after a genetic or chemical perturbation. Examining these populations alongside G0/G1, S, and G2/M distributions helps researchers characterize changes in genome replication and chromosome content.
The method connects fluorescence measurements with cellular DNA content, enabling researchers to examine genome replication, cell proliferation, and cell-cycle regulation. It is especially useful when genetic or chemical perturbations may change the distribution of cells across cycle phases or alter chromosome content. Flow-cytometric analysis provides these measurements across individual cells rather than only as a population average.