Nitrogen-15 Labeling
Nitrogen-15 Labeling - Related Videos
10:16
Design and Operation of a Continuous 13C and 15N Labeling Chamber for Uniform or Differential, Metabolic and Structural, Plant Isotope Labeling
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Cited by 39 •
2014
This method explains how to build and operate a continuous 13C and 15N isotope labeling chamber for uniform or differential plant tissue labeling. Representative results from metabolic and structural labeling of Andropogon gerardii are discussed.
08:44
Microplot Design and Plant and Soil Sample Preparation for 15Nitrogen Analysis
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Cited by 2 •
2020
A microplot design for 15N tracer research is described to accommodate multiple in-season plant and soil sampling events. Soil and plant sample collection and processing procedures, including grinding and weighing protocols, for 15N analysis are put forth.
14:44
A Protocol for the Identification of Protein-protein Interactions Based on 15N Metabolic Labeling, Immunoprecipitation, Quantitative Mass Spectrometry and Affinity Modulation
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Cited by 9 •
2012
We present a variation of the QUICK (QUantitative Immunoprecipitation Combined with Knockdown) approach that was introduced previously to distinguish between true and false protein-protein interactions. Our approach is based on 15N metabolic labeling, the modulation of affinities of protein-protein interactions by the presence/absence of ATP, immunoprecipitation, and quantitative mass spectrometry.
01:49
The Nitrogen Cycle
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2019
Nitrogen atoms, present in all proteins and DNA, are recycled between abiotic and biotic components of the ecosystem. However, the primary form of nitrogen on Earth is nitrogen gas, which cannot be used by most animals and plants. Thus, nitrogen gas must first be converted into a usable form by nitrogen-fixing bacteria before it can be cycled through other living organisms. The use of nitrogen-containing fertilizers and animal waste products in human agriculture has greatly influenced the...
08:04
A New Approach for the Comparative Analysis of Multiprotein Complexes Based on 15N Metabolic Labeling and Quantitative Mass Spectrometry
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Cited by 4 •
2014
The described comparative, quantitative proteomic approach aims at obtaining insights into the composition of multiprotein complexes under different conditions and is demonstrated by comparing genetically different strains. For quantitative analysis equal volumes of different fractions from a sucrose density gradient are mixed and analyzed by mass spectrometry.
FAQs
They identify incorporation by measuring changes associated with nitrogen-15 rather than simply detecting the molecule itself. Mass spectrometry resolves differences in molecular mass, while nuclear magnetic resonance detects characteristic signals from the labeled material. These measurements connect the presence of nitrogen-15 with specific molecular pools and provide evidence that nitrogen-containing compounds have been produced or modified during the experiment.
Nitrogen-15 labeling allows researchers to follow nitrogen movement without using radioactive tracers. Because the label can be detected through mass differences or nuclear magnetic resonance signals, experiments can relate molecular composition to cellular activity. This makes it useful for examining how biological systems build, replace, and redistribute nitrogen-containing compounds under defined physiological or environmental conditions.
The approach can be applied to protein synthesis and turnover, nucleic acid production, metabolic flux, and nutrient assimilation. These readouts address different aspects of biological activity: synthesis and turnover concern changes in molecular pools, metabolic flux concerns movement through biochemical pathways, and assimilation concerns the uptake and incorporation of available nitrogen into cellular material.
The label enters a system through nitrogen-containing nutrients or precursor compounds, so the chosen input determines which biological processes can be followed. Physiological and environmental conditions also matter because they influence how organisms process nitrogen. Comparing labeling under different conditions can therefore reveal changes in nutrient assimilation, molecular production, or nitrogen use rather than only showing label presence.
A typical workflow supplies cells or organisms with nitrogen-15 in a nutrient or precursor compound, allows biological processing to occur, and then measures labeled molecules using mass spectrometry or nuclear magnetic resonance. Researchers interpret the resulting mass differences or characteristic signals to determine how nitrogen entered molecular products and what cellular activities those products represent.
Researchers choose this approach when they need to connect nitrogen handling with molecular activity, including studies of protein turnover, nucleic acid production, metabolic flux, or nutrient assimilation. It is especially informative when conditions are varied to compare nitrogen processing across physiological or environmental states. The resulting measurements help explain how organisms transform available nitrogen into cellular material.