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Label-free quantification of membrane-ligand interactions using backscattering interferometry
by
Baksh, Michael M
, Bornhop, Darryl J
, Mileni, Mauro
, Finn, M G
, Kussrow, Amanda K
in
631/61/32
/ Agriculture
/ Amidohydrolases - metabolism
/ Binding sites
/ Bioinformatics
/ Biological and medical sciences
/ Biomedical and Life Sciences
/ Biomedical Engineering/Biotechnology
/ Biomedicine
/ Biotechnology
/ Cellular biology
/ Cholera Toxin - metabolism
/ Diverse techniques
/ Fundamental and applied biological sciences. Psychology
/ G(M1) Ganglioside - metabolism
/ Humans
/ Interferometry
/ Interferometry - methods
/ Kinetics
/ letter
/ Life Sciences
/ Ligands
/ Ligands (Biochemistry)
/ Membrane proteins
/ Membrane Proteins - metabolism
/ Membranes
/ Methods
/ Microscopy, Fluorescence
/ Molecular and cellular biology
/ Properties
/ Protein Binding
/ Proteins
/ Receptors, CXCR4 - metabolism
/ Receptors, GABA-B - metabolism
/ Refractometry - methods
2011
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Label-free quantification of membrane-ligand interactions using backscattering interferometry
by
Baksh, Michael M
, Bornhop, Darryl J
, Mileni, Mauro
, Finn, M G
, Kussrow, Amanda K
in
631/61/32
/ Agriculture
/ Amidohydrolases - metabolism
/ Binding sites
/ Bioinformatics
/ Biological and medical sciences
/ Biomedical and Life Sciences
/ Biomedical Engineering/Biotechnology
/ Biomedicine
/ Biotechnology
/ Cellular biology
/ Cholera Toxin - metabolism
/ Diverse techniques
/ Fundamental and applied biological sciences. Psychology
/ G(M1) Ganglioside - metabolism
/ Humans
/ Interferometry
/ Interferometry - methods
/ Kinetics
/ letter
/ Life Sciences
/ Ligands
/ Ligands (Biochemistry)
/ Membrane proteins
/ Membrane Proteins - metabolism
/ Membranes
/ Methods
/ Microscopy, Fluorescence
/ Molecular and cellular biology
/ Properties
/ Protein Binding
/ Proteins
/ Receptors, CXCR4 - metabolism
/ Receptors, GABA-B - metabolism
/ Refractometry - methods
2011
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Label-free quantification of membrane-ligand interactions using backscattering interferometry
by
Baksh, Michael M
, Bornhop, Darryl J
, Mileni, Mauro
, Finn, M G
, Kussrow, Amanda K
in
631/61/32
/ Agriculture
/ Amidohydrolases - metabolism
/ Binding sites
/ Bioinformatics
/ Biological and medical sciences
/ Biomedical and Life Sciences
/ Biomedical Engineering/Biotechnology
/ Biomedicine
/ Biotechnology
/ Cellular biology
/ Cholera Toxin - metabolism
/ Diverse techniques
/ Fundamental and applied biological sciences. Psychology
/ G(M1) Ganglioside - metabolism
/ Humans
/ Interferometry
/ Interferometry - methods
/ Kinetics
/ letter
/ Life Sciences
/ Ligands
/ Ligands (Biochemistry)
/ Membrane proteins
/ Membrane Proteins - metabolism
/ Membranes
/ Methods
/ Microscopy, Fluorescence
/ Molecular and cellular biology
/ Properties
/ Protein Binding
/ Proteins
/ Receptors, CXCR4 - metabolism
/ Receptors, GABA-B - metabolism
/ Refractometry - methods
2011
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Label-free quantification of membrane-ligand interactions using backscattering interferometry
Journal Article
Label-free quantification of membrane-ligand interactions using backscattering interferometry
2011
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Overview
Methods to measure affinities of membrane proteins and soluble ligands are cumbersome and often rely on truncations or other modifications of the membrane protein or ligand. Baksh
et al
. show that backscattering interferometry is a sensitive and accurate technology for the label-free quantification of ligand–membrane receptor interactions.
Although membrane proteins are ubiquitous within all living organisms and represent the majority of drug targets, a general method for direct, label-free measurement of ligand binding to native membranes has not been reported. Here we show that backscattering interferometry (BSI) can accurately quantify ligand-receptor binding affinities in a variety of membrane environments. By detecting minute changes in the refractive index of a solution, BSI allows binding interactions of proteins with their ligands to be measured at picomolar concentrations. Equilibrium binding constants in the micromolar to picomolar range were obtained for small- and large-molecule interactions in both synthetic and cell-derived membranes without the use of labels or supporting substrates. The simple and low-cost hardware, high sensitivity and label-free nature of BSI should make it readily applicable to the study of many membrane-associated proteins of biochemical and pharmacological interest.
Publisher
Nature Publishing Group US,Nature Publishing Group
Subject
/ Amidohydrolases - metabolism
/ Biological and medical sciences
/ Biomedical and Life Sciences
/ Biomedical Engineering/Biotechnology
/ Fundamental and applied biological sciences. Psychology
/ G(M1) Ganglioside - metabolism
/ Humans
/ Kinetics
/ letter
/ Ligands
/ Membrane Proteins - metabolism
/ Methods
/ Molecular and cellular biology
/ Proteins
/ Receptors, CXCR4 - metabolism
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