Biacore X100
Biacore X100 - Related Videos
15:27
Determination of High-affinity Antibody-antigen Binding Kinetics Using Four Biosensor Platforms
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Cited by 30 •
2017
We describe here protocols for the measurement of antibody-antigen binding affinity and kinetics using four commonly used biosensor platforms.
12:17
Initiating Differentiation in Immortalized Multipotent Otic Progenitor Cells
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Cited by 10 •
2016
The current protocols to maintain immortalized multipotent otic progenitor (iMOP) cells and otic differentiation are described. Culture conditions and molecular markers that indicate differentiation into sensory epithelia and spiral ganglion neurons (SGN) are highlighted.
09:34
Study of Endoplasmic Reticulum and Mitochondria Interactions by In Situ Proximity Ligation Assay in Fixed Cells
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Cited by 59 •
2016
Here, we describe a procedure to visualize and quantify with high sensitivity the endogenous interactions between the endoplasmic reticulum and mitochondria in fixed cells. The protocol features an optimized in situ proximity ligation assay targeting the inositol 1,4,5-triphosphate receptor/glucose-regulated protein 75/voltage-dependent anion channel/cyclophilin D complex at the mitochondria-associated membrane interface.
08:09
Vasodilation of Isolated Vessels and the Isolation of the Extracellular Matrix of Tight-skin Mice
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Cited by 1 •
2017
We describe the isolation of cardiac extracellular matrix from C57Bl/6J control mice, tight-skin mice, and tight-skin mice treated with the IRF5 inhibitory peptide. We also describe the vasodilation studies on the isolated vessels from C57Bl/6J, tight-skin mice and tight-skin mice treated with the IRF5 inhibitory peptide.
08:37
In Situ MHC-tetramer Staining and Quantitative Analysis to Determine the Location, Abundance, and Phenotype of Antigen-specific CD8 T Cells in Tissues
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Cited by 11 •
2017
Here, we describe a method that combines in situ MHC-tetramer staining with immunohistochemistry to determine localization, phenotype, and quantity of antigen-specific T cells in tissues. This protocol is used to determine the spatial and phenotypic characteristics of antigen-specific CD8 T cells relative to other cell type and structures in tissues.
FAQs
Biacore X100 tracks changes at the sensor surface as molecules bind and leave. When the flowing binding partner associates with the immobilized partner, the local refractive index changes and produces a response. During dissociation, the signal changes as bound molecules depart. The resulting sensorgram therefore contains time-resolved information rather than only an endpoint measurement.
Association and dissociation portions of a sensorgram are interpreted together to characterize an interaction. Association behavior reports how the flowing molecule binds to the surface-bound partner, whereas dissociation behavior follows what happens after the binding phase. Analysis of these phases provides binding kinetics and affinity, allowing comparisons among biomolecular interactions.
Because Biacore X100 does not require labels, the interacting molecules can be monitored directly through their surface response. The readout comes from the binding event and its effect on the refractive index rather than from a separate labeling signal. This capability supports real-time observation of association and dissociation during studies of molecular recognition.
A typical measurement begins with a sensor chip carrying one binding partner. The other partner is prepared in solution and passed across the surface under flow. The system records the response over time, generating a sensorgram for analysis. This workflow can be applied to proteins, nucleic acids, antibodies, or small molecules, depending on the interaction being studied.
The platform can examine interactions involving proteins, nucleic acids, antibodies, and small molecules. Researchers can use the same real-time binding framework to investigate molecular recognition while adapting the binding partners to the biological question. This breadth makes it useful for studying diverse interaction classes rather than a single type of biomolecule.
In biology, Biacore X100 measurements support target validation, assay development, and therapeutic characterization. They can also clarify molecular recognition by showing how strongly and how quickly partners associate and dissociate. These outputs help connect a measured interaction with the biological or development question that motivated the experiment.