Iodide-yfp-gjic Assay
Iodide-yfp-gjic Assay - Related Videos
04:31
Differential Nuclear Staining Assay: An Assay to Determine Mitocan Cytotoxicity by Propidium Iodide and Hoechst Double Staining
0 Views •
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.
09:09
Annexin V and Propidium Iodide Labeling
0 Views •
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
0 Views •
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.
10:03
Multiphoton Microscopy of Cleared Mouse Brain Expressing YFP
0 Views •
Cited by 11 •
2012
Multiphoton microscopy of whole mouse organs is possible by optically clearing the organ before imaging, but not all protocols preserve the fluorescent signal of fluorescent proteins. Using an optical clearing method with ethanol-based dehydration and benzyl alcohol:benzyl benzoate clearing, we show high-resolution multiphoton images of whole mouse brain expressing YFP.
09:47
An Iodide-Yellow Fluorescent Protein-Gap Junction-Intercellular Communication Assay
0 Views •
Cited by 3 •
2019
Here, we present a protocol for a novel gap junction intercellular communication assay designed for the high-throughput screening of gap junction-modulating chemicals for drug discovery and toxicological assessment.
FAQs
Iodide acts as a quencher for the yellow fluorescent protein signal. After exposure, its movement from one cell into neighboring cells depends on functional gap junction channels, so fluorescence decreases as iodide reaches YFP-expressing cells. Monitoring this loss over time converts molecular transfer between cells into a measurable fluorescence-based indicator of junctional communication.
The rate of fluorescence decline reflects how quickly iodide moves through communicating cells, whereas the extent of signal loss reflects how much transfer occurs during the measurement. Considering both values provides more information than a single endpoint: one describes the dynamics of communication, and the other indicates the overall transfer detected under the tested conditions.
Changes in the fluorescence response can indicate altered activity of the gap junction pathways formed by connexins. A different rate or extent of YFP quenching may therefore reflect changes in cellular connectivity or channel function. This makes the assay useful for examining how experimental conditions or candidate treatments modify intercellular communication.
Cells must first express an iodide-sensitive yellow fluorescent protein, after which iodide is introduced to the cell system. Fluorescence is then monitored as iodide reaches neighboring cells through gap junction channels. The resulting time-dependent signal loss is quantified by assessing both how rapidly fluorescence decreases and how much it declines.
Researchers can compare fluorescence responses from treated and untreated cell systems to determine whether a candidate treatment changes gap junction communication. Differences in the speed or magnitude of YFP quenching provide a quantitative comparison of iodide transfer. This approach helps identify conditions that enhance, reduce, or otherwise modify cellular connectivity.
The method links a visible fluorescence change to the direct exchange of a small molecule between neighboring cells. Because the signal can be quantified through its rate and extent of decline, it provides an experimental readout of how well cells communicate. This supports studies of connexin function, intercellular connectivity, and condition-dependent changes in tissue-related cell behavior.