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Biacore spr experiments
(A) Binding isotherms of the IL-13 agonists displayed on the surface of yeast. Yeast-displayed IL-13 agonists were incubated with the indicated concentrations of biotinylated IL-13Rα1 ECD, stained with Alexa-647-coupled Streptavidin, and assessed via flow cytometry. Sigmoidal curves were fitted using Prism software (GraphPad). (B) Normalized KD binding affinities of the recombinant IL-13 agonists determined via surface <t>plasmon</t> <t>resonance</t> <t>(SPR).</t> The IL-13 KD was fixed to one and other agonists were normalized accordingly. (C) Summary of the amino acids mutated in the IL-13 agonists as well as the kon, koff and KD binding parameters values obtained from SPR experiments.
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Biacore surface plasmon resonance-based solution affinity assay
(A) Binding isotherms of the IL-13 agonists displayed on the surface of yeast. Yeast-displayed IL-13 agonists were incubated with the indicated concentrations of biotinylated IL-13Rα1 ECD, stained with Alexa-647-coupled Streptavidin, and assessed via flow cytometry. Sigmoidal curves were fitted using Prism software (GraphPad). (B) Normalized KD binding affinities of the recombinant IL-13 agonists determined via surface <t>plasmon</t> <t>resonance</t> <t>(SPR).</t> The IL-13 KD was fixed to one and other agonists were normalized accordingly. (C) Summary of the amino acids mutated in the IL-13 agonists as well as the kon, koff and KD binding parameters values obtained from SPR experiments.
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Federation of European Neuroscience Societies surface plasmon resonance
(A) Binding isotherms of the IL-13 agonists displayed on the surface of yeast. Yeast-displayed IL-13 agonists were incubated with the indicated concentrations of biotinylated IL-13Rα1 ECD, stained with Alexa-647-coupled Streptavidin, and assessed via flow cytometry. Sigmoidal curves were fitted using Prism software (GraphPad). (B) Normalized KD binding affinities of the recombinant IL-13 agonists determined via surface <t>plasmon</t> <t>resonance</t> <t>(SPR).</t> The IL-13 KD was fixed to one and other agonists were normalized accordingly. (C) Summary of the amino acids mutated in the IL-13 agonists as well as the kon, koff and KD binding parameters values obtained from SPR experiments.
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Verlag GmbH surface plasmon resonances (lsprs)
(A) Binding isotherms of the IL-13 agonists displayed on the surface of yeast. Yeast-displayed IL-13 agonists were incubated with the indicated concentrations of biotinylated IL-13Rα1 ECD, stained with Alexa-647-coupled Streptavidin, and assessed via flow cytometry. Sigmoidal curves were fitted using Prism software (GraphPad). (B) Normalized KD binding affinities of the recombinant IL-13 agonists determined via surface <t>plasmon</t> <t>resonance</t> <t>(SPR).</t> The IL-13 KD was fixed to one and other agonists were normalized accordingly. (C) Summary of the amino acids mutated in the IL-13 agonists as well as the kon, koff and KD binding parameters values obtained from SPR experiments.
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Chemie GmbH surface plasmon resonance (spr)
(A) Binding isotherms of the IL-13 agonists displayed on the surface of yeast. Yeast-displayed IL-13 agonists were incubated with the indicated concentrations of biotinylated IL-13Rα1 ECD, stained with Alexa-647-coupled Streptavidin, and assessed via flow cytometry. Sigmoidal curves were fitted using Prism software (GraphPad). (B) Normalized KD binding affinities of the recombinant IL-13 agonists determined via surface <t>plasmon</t> <t>resonance</t> <t>(SPR).</t> The IL-13 KD was fixed to one and other agonists were normalized accordingly. (C) Summary of the amino acids mutated in the IL-13 agonists as well as the kon, koff and KD binding parameters values obtained from SPR experiments.
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Danaher Inc biacore 3000 instrument
(A) Binding isotherms of the IL-13 agonists displayed on the surface of yeast. Yeast-displayed IL-13 agonists were incubated with the indicated concentrations of biotinylated IL-13Rα1 ECD, stained with Alexa-647-coupled Streptavidin, and assessed via flow cytometry. Sigmoidal curves were fitted using Prism software (GraphPad). (B) Normalized KD binding affinities of the recombinant IL-13 agonists determined via surface <t>plasmon</t> <t>resonance</t> <t>(SPR).</t> The IL-13 KD was fixed to one and other agonists were normalized accordingly. (C) Summary of the amino acids mutated in the IL-13 agonists as well as the kon, koff and KD binding parameters values obtained from SPR experiments.
Biacore 3000 Instrument, supplied by Danaher Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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(A) Binding isotherms of the IL-13 agonists displayed on the surface of yeast. Yeast-displayed IL-13 agonists were incubated with the indicated concentrations of biotinylated IL-13Rα1 ECD, stained with Alexa-647-coupled Streptavidin, and assessed via flow cytometry. Sigmoidal curves were fitted using Prism software (GraphPad). (B) Normalized KD binding affinities of the recombinant IL-13 agonists determined via surface plasmon resonance (SPR). The IL-13 KD was fixed to one and other agonists were normalized accordingly. (C) Summary of the amino acids mutated in the IL-13 agonists as well as the kon, koff and KD binding parameters values obtained from SPR experiments.

Journal: Science signaling

Article Title: Instructive roles for agonist binding parameters in determining the functional bandwidth of cytokine receptor signaling

doi: 10.1126/scisignal.aab2677

Figure Lengend Snippet: (A) Binding isotherms of the IL-13 agonists displayed on the surface of yeast. Yeast-displayed IL-13 agonists were incubated with the indicated concentrations of biotinylated IL-13Rα1 ECD, stained with Alexa-647-coupled Streptavidin, and assessed via flow cytometry. Sigmoidal curves were fitted using Prism software (GraphPad). (B) Normalized KD binding affinities of the recombinant IL-13 agonists determined via surface plasmon resonance (SPR). The IL-13 KD was fixed to one and other agonists were normalized accordingly. (C) Summary of the amino acids mutated in the IL-13 agonists as well as the kon, koff and KD binding parameters values obtained from SPR experiments.

Article Snippet: Surface plasmon resonance SPR experiments were conducted on a Biacore T100 instrument using a Biacore SA sensor chip (GE Healthcare).

Techniques: Binding Assay, Incubation, Staining, Flow Cytometry, Software, Recombinant, SPR Assay