Radiolabeling With 3h-inositol
Radiolabeling With 3h-inositol - Related Videos
09:01
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Here we describe strong anion exchange high-performance liquid chromatography of [3H]-myo-inositol-labeled seedlings which is a highly sensitive method to detect and quantify inositol polyphosphates in plants.
03:28
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In this video, we demonstrate the intracellular radiolabeling of T helper cells using radioactive isotope-conjugated monoclonal antibodies for non-invasive positron emission tomography imaging.
10:01
Measurement of Carbon Dioxide Production from Radiolabeled Substrates in Drosophila melanogaster
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03:35
Assessing Viral Replication Capacity Using a Radiolabeled Reverse Transcriptase Assay
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2026
Source: Claiborne, D. T. et al. A Restriction Enzyme Based Cloning Method to Assess the In vitro Replication Capacity of HIV-1 Subtype C Gag-MJ4 Chimeric Viruses. J. Vis. Exp. (2014)This video demonstrates the assessment of viral replication capacity using a radiolabeled reverse transcriptase assay. Culture supernatants containing progeny virions are added to a reverse transcription mixture containing a radiolabeled nucleotide. The incorporated signal reflects viral output and replication...
10:52
Radiolabeling and Quantification of Cellular Levels of Phosphoinositides by High Performance Liquid Chromatography-coupled Flow Scintillation
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Cited by 21 •
2016
Phosphoinositides are signaling lipids whose relative abundance rapidly changes in response to various stimuli. This article describes a method to measure the abundance of phosphoinositides by metabolically labeling cells with 3H-myo-inositol, followed by extraction and deacylation. Extracted glycero-inositides are then separated by high-performance liquid chromatography and quantified by flow scintillation.
FAQs
Cells incorporate [3H]inositol into phosphoinositides, placing the radioactive label within membrane-associated inositol lipids. When a receptor stimulus activates phospholipase C, the label can appear in newly generated inositol phosphates derived from those lipids. Tracking this movement connects receptor activation with downstream biochemical changes and helps reveal how cellular signaling alters inositol-containing metabolites.
Phospholipase C provides the mechanistic link between receptor stimulation and labeled product formation. After receptor activation, it produces inositol phosphates from labeled phosphoinositides in the membrane. Measuring those products therefore gives a biochemical readout of receptor-linked signaling, rather than merely indicating that radioactive inositol was present in the cells.
Chromatographic separation distinguishes the radioactive products before their amount is assessed, while scintillation counting quantifies their radioactivity. Using both steps makes it possible to associate measured signal with separated inositol-containing products instead of treating all cellular radioactivity as one pool. This supports analysis of phosphoinositide metabolism after receptor stimulation.
A typical workflow allows cells to incorporate [3H]inositol into phosphoinositides, applies a receptor stimulus, and then examines the labeled inositol-containing products generated during the response. Researchers separate these radioactive products, often chromatographically, and quantify them by scintillation counting. Comparing the measured products helps assess changes in phosphoinositide metabolism and signaling.
The measured products can indicate changes in inositol-lipid synthesis, turnover, and signaling activity. They also help characterize receptor-linked pathways and enzyme activity by showing how stimulation changes the production of labeled inositol phosphates. Consequently, the technique can connect an extracellular stimulus with a quantifiable intracellular biochemical response.
Radiolabeling with 3H-inositol is useful when researchers need to examine how receptors influence phosphoinositide metabolism or intracellular second-messenger production. It can support studies of signaling pathways, enzyme activity, and changes in cellular responses following stimulation. The radioactive label provides a sensitive way to follow inositol-containing compounds through these biochemical processes.