Lc-ms/ms Analysis
Lc-ms/ms Analysis - Related Videos
11:16
Discovering Protein Interactions and Characterizing Protein Function Using HaloTag Technology
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Cited by 11 •
2014
HaloTag technology is a multifunctional technology which has shown significant success in isolation of both small and large protein complexes from mammalian cells. Here we highlight the advantages of this technology compared to existing alternatives and demonstrate its utility to study numerous aspects of protein function inside eukaryotic cells.
10:49
Printing Thermoresponsive Reverse Molds for the Creation of Patterned Two-component Hydrogels for 3D Cell Culture
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Cited by 59 •
2013
A bioprinter was used to create patterned hydrogels based on a sacrificial mold. The poloxamer mold was backfilled with a second hydrogel and then eluted, leaving voids which were filled with a third hydrogel. This method uses fast elution and good printability of poloxamer to generate complex architectures from biopolymers.
06:49
Tracking Drosophila Larval Behavior in Response to Optogenetic Stimulation of Olfactory Neurons
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Cited by 8 •
2018
This protocol analyzes navigational behavior of Drosophila larva in response to simultaneous optogenetic stimulation of its olfactory neurons. Light of 630 nm wavelength is used to activate individual olfactory neurons expressing a red-shifted channel rhodopsin. Larval movement is simultaneously tracked, digitally recorded, and analyzed using custom-written software.
10:22
Basophil Activation Test for Investigation of IgE-Mediated Mechanisms in Drug Hypersensitivity
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Cited by 14 •
2011
Basophil activation test is a potent tool for the detection of IgE-dependent allergies in vitro. Here, an optimized protocol for basophil activation test is used to investigate drug hypersensitivity. A method for the efficient production of covalent drug-protein conjugates and their physicochemical characterization is described.
15:04
A Neuronal and Astrocyte Co-Culture Assay for High Content Analysis of Neurotoxicity
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Cited by 46 •
2009
This article describes a novel protocol and reagent set designed for sensitive measurement of neurotoxic effects of compounds and treatments on co-cultures of neurons and astrocytes using high content analysis. Results demonstrate that high content analysis represents an exciting novel technology for neurotoxicity assessment.
FAQs
Liquid chromatography separates compounds before mass analysis according to how they partition between the mobile and stationary phases. This reduces the number of molecules entering the mass spectrometer at the same time, helping distinguish components in complex biological samples. The resulting separation supports more reliable identification and quantification of metabolites, lipids, peptides, proteins, and other biomolecules.
A precursor ion is selected according to its mass-to-charge ratio and then fragmented inside the instrument. The resulting product ions provide a characteristic pattern that supports molecular identification. Measuring these fragments also contributes to selective quantification, because the analysis evaluates both the original ion and its fragmentation products rather than relying on mass alone.
Targeted analysis focuses on selected molecules and measures them specifically, making it useful when researchers want to assess defined metabolites, lipids, peptides, or other biomolecules. Untargeted analysis surveys molecular features more broadly to help characterize biological changes and discover candidates for further study. The choice depends on whether the investigation begins with defined targets or exploratory biological questions.
LC-MS/MS analysis can be applied to diverse biomolecular classes, including metabolites, lipids, peptides, proteins, and other molecules present in biological samples. This breadth allows one analytical platform to support investigations at several biological levels, from molecular products and lipid composition to peptide or protein changes associated with physiology, disease, or treatment.
The workflow begins with liquid chromatographic separation of compounds in a biological sample. The separated molecules are then ionized, and the mass spectrometer selects precursor ions by mass-to-charge ratio. Those ions are fragmented, and the instrument measures the resulting product ions. Researchers use the combined separation and spectral information to identify and quantify molecular components.
Researchers use this approach to characterize molecular pathways, compare physiological and disease states, identify biomarkers, and measure treatment-related molecular changes. Its high sensitivity and selectivity are valuable when biological samples contain many chemically different components. Results can reveal which molecules change between conditions and can connect those changes with pathways, disease processes, or responses to treatment.