Lc-ms/ms Assay
Lc-ms/ms Assay - Related Videos
11:16
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Cited by 11 •
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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
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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.
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Cited by 8 •
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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.
08:12
The TUNEL Assay
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2023
One of the hallmarks of apoptosis is the nuclear DNA fragmentation by nucleases. These enzymes are activated by caspases, the family of proteins that execute the cell death program. TUNEL assay is a method that takes advantage of this feature to detect apoptotic cells. In this assay, an enzyme called terminal deoxynucleotidyl transferase catalyzes the addition of dUTP nucleotides to the free 3’ ends of fragmented DNA. By using dUTPs that are labeled with chemical tags that can produce...
10:22
Basophil Activation Test for Investigation of IgE-Mediated Mechanisms in Drug Hypersensitivity
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Cited by 14 •
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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.
FAQs
Separation depends on how each analyte interacts with the stationary phase and mobile phase. These interactions determine when compounds are resolved from one another before mass analysis. Better resolution helps distinguish analytes in complex samples, supporting more reliable identification and quantification of metabolites, peptides, lipids, drugs, and other biochemical compounds.
The instrument first selects a precursor ion associated with an analyte, then measures product ions formed after fragmentation. This sequential selection provides characteristic mass information beyond the original ion alone. Using both ion types helps distinguish chemically related compounds and supports confident identification while the measured response contributes to quantification.
Calibration standards establish the relationship between analyte measurement and quantity, allowing the assay to determine concentrations in unknown samples. Internal standards provide a reference during measurement, helping support accuracy when analytes are examined in biological matrices. Together, these materials make quantitative results more dependable across complex sample types.
A typical workflow begins by resolving compounds with liquid chromatography, followed by ionization for mass analysis. The system selects a precursor ion, measures characteristic product ions generated by fragmentation, and compares measurements with calibration standards. Internal standards are also used to support accurate quantification, particularly when samples contain complex biological material.
The assay can measure a broad range of biochemical targets, including metabolites, peptides, lipids, drugs, and other biomolecules. This breadth makes it useful when biological samples contain chemically different analytes that require both chromatographic separation and mass-based identification. Quantitative measurements can then support biochemical investigations rather than limiting analysis to a single compound class.
In biochemistry, researchers apply LC-MS/MS assays to metabolic research, biomarker validation, pharmaceutical analysis, and clinical investigation. The method can provide measurements from complex biological samples, helping evaluate metabolites, peptides, lipids, drugs, and other biomolecules. Its combination of identification and quantification is especially relevant when studies require measured analyte levels rather than qualitative observation alone.