In-situ photopolymerized C4-functionalized organosilicon monoliths for reversed-phase protein separation in nano-liquid chromatography

Talanta ◽  
2019 ◽  
Vol 198 ◽  
pp. 330-336 ◽  
Author(s):  
Çiğdem Kip ◽  
Siyao Liu ◽  
Xinyue Fu ◽  
Ali Tuncel ◽  
Michael Lämmerhofer
Keyword(s):  
Liquid Chromatography ◽  
Protein Separation ◽  
Reversed Phase ◽  
Phase Protein ◽  
Nano Liquid Chromatography

scholarly journals A Fast and Accurate Method to Identify and Quantify Enzymes in Brush-Border Membranes: In Situ Hydrolysis Followed by Nano LC-MS/MS

2020 ◽  
Vol 3 (1) ◽  
pp. 15 ◽  
Author(s):  
Antonio Brun ◽  
Melisa E. Magallanes ◽  
Carlos Martínez del Rio ◽  
Gregory A. Barrett-Wilt ◽  
William H. Karasov ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Brush Border ◽  
Brush Border Membrane ◽  
Accurate Method ◽  
Simple Method ◽  
Brush Border Membranes ◽  
Nano Liquid Chromatography ◽  
Orange Color

A simple method for the identification of brush-border membrane α-glucosidases is described. The proteins were first solubilized and separated in a gel under native, non-denaturing, conditions. The gel was then incubated in substrate solutions (maltose or sucrose), and the product (glucose) exposed in situ by the oxidation of o-dianisidine, which yields a brown-orange color. Nano-liquid chromatography coupled to mass spectrometry analyses of proteins (nano LC-MS/MS) present in the colored bands excised from the gels, was used to confirm the presence of the enzymes. The stain is inexpensive and the procedure permits testing several substrates in the same gel. Once enzymes are identified, their abundance, relative to that of other proteins in the brush border, can be semi-quantified using nano LC-MS/MS.

Liquid chromatography with amperometric reaction detection involving electrogenerated reagents: applications with in-situ generated bromine.

1980 ◽  
Vol 26 (10) ◽  
pp. 1484-1491 ◽  
Author(s):  
W P King ◽  
P T Kissinger
Keyword(s):  
Liquid Chromatography ◽  
Reversed Phase ◽  
Reagent Concentration ◽  
Constant Rate ◽  
On Line ◽  
Electrochemical Control ◽  
Phase Liquid ◽  
Direct Generation ◽  
Liquid Chromatographic

Abstract We describe the use of electrogenerated reactants for continuous on-line reaction detection with thin-layer hydrodynamic amperometry. The reagent is introduced into the liquid-chromatographic column effluent at a constant rate by using controlled-current electrochemistry. After the effluent passes through a short reaction coil, the reagent concentration is monitored at the detector. Reaction of eluted compounds with bromine is signalled by changes in the current detected. The direct electrochemical control of the reagent concentration allows changes to be made, even during the course of obtaining a chromatogram. Depending on the specific reagent or reaction, the reagent is supplied either by addition of a second stream or by direct generation in the mobile phase. The latter configuration provides sufficient baseline stability to permit detection of the uptake of as little as 10 pmol of reagent bromine. The technique has been used to detect nanograms of underivatized fatty acids, prostaglandins, and phenols after separation by reversed-phase liquid chromatography.

Perfusion reversed-phase high-performance liquid chromatography for protein separation from detergent-containing solutions: An alternative to gel-based approaches

2012 ◽  
Vol 424 (2) ◽  
pp. 97-107 ◽  
Author(s):  
Jan Gorka ◽  
Marion Rohmer ◽  
Sandra Bornemann ◽  
Dimitrios G. Papasotiriou ◽  
Dominic Baeumlisberger ◽  
...  
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Protein Separation ◽  
High Performance ◽  
Reversed Phase
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