Ionophores

Ionophores are small, typically lipid-soluble molecules that transport specific ions across biological membranes, altering the electrical and chemical gradients cells use for signaling, metabolism, and homeostasis. They work by either forming membrane-spanning channels or binding ions and carrying them through the hydrophobic lipid bilayer; their activity depends on ion selectivity, concentration gradients, and membrane potential. In biology, ionophores help researchers manipulate intracellular calcium, potassium, sodium, or proton levels to investigate membrane transport, mitochondrial function, secretion, and cell death. Naturally occurring and synthetic ionophores also have applications as antimicrobial or antiparasitic agents, while their ability to disrupt ion balance makes dose and cellular context critical.

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Research

JoVE EoE - Assay Techniques

Hemolysis Assay: An In Vitro Method to Measure Calcium Ionophore-Induced Lysis in Human Erythrocytes

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2025

This video describes an in vitro assay to determine calcium-ionophore-induced hemolysis or lysis of red blood cells in a human sample. The assay measures the percentage of hemolysis by quantifying hemoglobin released from ruptured erythrocytes.

Induction of Eryptosis in Red Blood Cells Using a Calcium Ionophore

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Cited by 21 •

2020

A protocol for the induction of eryptosis, programmed cell death in erythrocytes, using the calcium ionophore, ionomycin, is provided. Successful eryptosis is evaluated by monitoring the localization phosphatidylserine in the membrane outer leaflet. Factors affecting the success of the protocol have been examined and optimal conditions provided.

An In Vivo Model to Study Shear-Dependent Bacterial Adhesion

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2026

Source: Claes, J., et al, In Vitro and In Vivo Model to Study Bacterial Adhesion to the Vessel Wall Under Flow Conditions. J. Vis. Exp. (2015).This video demonstrates an in vivo method for analyzing shear-dependent bacterial adhesion in a live mouse model. Following surgical preparation and catheterization, the mesenteric vasculature is exposed for real-time imaging under a fluorescence microscope. Perfusion with an ionophore and fluorescently labeled bacteria reveals how shear stress triggers...

Annexin V Binding Assay: A Fluorescence-Based Technique to Identify Apoptotic Erythrocytes via Phosphatidylserine Labeling

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2025

In this video, we demonstrate the Annexin V binding assay to measure the degree of eryptosis, erythrocyte programmed cell death, using fluorescent Annexin V staining. Apoptotic erythrocytes demonstrate phosphatidylserine translocation from the inner to the outer leaflet of the cell membrane, causing the fluorophore-tagged Annexin V to bind to the translocated phosphatidylserine, thereby emitting a fluorescent signal.

Measuring Mast Cell Exocytosis via Neuropeptide Y Monomeric Red Fluorescent Protein Release

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2025

This video demonstrates mast cell exocytosis induced by an activating reagent that includes a calcium ionophore, initiating the release of secretory granules containing the fluorescent reporter protein NPY-mRFP. Subsequently, we quantitatively assess the exocytosis by measuring the released NPY-mRFP using a fluorescence plate reader.

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