Reversed-phase Lc-ms
Reversed-phase Lc-ms - Related Videos
03:31
Reversed-Phase High-Performance Liquid Chromatography: A Robust Method for Quantitation of Fluorescently Labeled and Derivatized Sialic Acids Isolated from Mouse Liver
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2025
In this video, we demonstrate the detection of sialic acids, N-acetylneuraminic acid and N-glycolylneuraminic acid, using high-performance liquid chromatography, HPLC, following derivatization with a suitable fluorescent labeling reagent to aid in fluorescence detection.
12:22
The Use of Reverse Phase Protein Arrays (RPPA) to Explore Protein Expression Variation within Individual Renal Cell Cancers
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Cited by 12 •
2013
RPPA enables the protein expression of hundreds of samples, printed on nitrocellulose slides to be interrogated simultaneously, using fluorescently labelled antibodies. This technique has been applied to study the effect of drug treatment heterogeneity within clear cell renal carcinoma.
10:49
Printing Thermoresponsive Reverse Molds for the Creation of Patterned Two-component Hydrogels for 3D Cell Culture
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Cited by 60 •
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.
06:32
Reverse Phase Protein Arrays Based Protein Expression Analysis: A Procedure for Simultaneous Quantification of Expression of Multiple Proteins from Cell Lysate
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2023
This video demonstrates the methodology for performing Reverse Phase Protein Arrays (RPPA) to study protein expression patterns from cell lysate. In RPPA, the lysate containing a mixture of proteins is printed on nitrocellulose slides and proteins of interest are analyzed using fluorescently labeled antibodies.
02:33
Phase Transitions: Sublimation and Deposition
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2020
Some solids can transition directly into the gaseous state, bypassing the liquid state, via a process known as sublimation. At room temperature and standard pressure, a piece of dry ice (solid CO2) sublimes, appearing to gradually disappear without ever forming any liquid. Snow and ice sublimate at temperatures below the melting point of water, a slow process that may be accelerated by winds and the reduced atmospheric pressures at high altitudes. When solid iodine is warmed, the solid sublimes...
FAQs
In the column, analytes distribute between the nonpolar stationary phase and polar mobile phase. Increasing organic-solvent content changes that balance, allowing compounds with different hydrophobicities to leave the column at different times. This time-based separation reduces mixture complexity before detection and helps assign mass-spectrometric signals to chromatographic peaks.
Electrospray ionization converts compounds emerging from the liquid stream into gas-phase ions, which makes them suitable for mass analysis. The instrument then measures their mass-to-charge ratios rather than relying only on chromatographic retention. This conversion connects the separated biochemical components with molecular-mass information for identification or quantification.
Retention behavior and mass-to-charge information provide complementary evidence. A compound’s position in the chromatogram reflects its interaction with the reversed-phase system, while its mass-spectrometric signal supplies molecular-mass information. Using both dimensions is especially valuable for complex biochemical samples, where separation alone or mass information alone may be less informative.
An analysis first passes the sample through a reversed-phase liquid-chromatography system, using an increasing organic-solvent gradient to elute components according to hydrophobicity. As each component exits, electrospray ionization produces gas-phase ions, and the mass spectrometer records mass-to-charge information. The resulting chromatographic and mass data support compound characterization.
The technique can be applied to peptides, proteins, metabolites, lipids, and pharmaceutical compounds. That breadth makes it useful when a biochemical investigation must examine chemically diverse molecules rather than a single purified class. These target categories connect the method with protein characterization, metabolic profiling, pharmaceutical analysis, and broader biochemical measurements.
It supports biomarker studies, protein characterization, metabolic profiling, and quantitative research. In these settings, chromatographic separation helps organize components from complex samples, while mass spectrometry contributes molecular-mass information and sensitive detection. Together, these outputs support characterization of biochemical composition and measurement of analytes across diverse research workflows.