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Protocol for Notch-ligand Binding Assays Using Dynabeads
Shogo Sawaguchi1, Mitsutaka Ogawa1, Tetsuya Okajima1
1Department of Biochemistry II, Nagoya University Graduate School of Medicine, Nagoya, Japan.
Abstract:
This protocol describes how to measure interaction between Notch receptors and their ligands by cell-based assay using Dynabeads. We have used the protocol to determine binding capacity between Notch1-transfected HEK293T cells and ligand-coated Dynabeads. Expression of Eogt in Notch1-expressing cells promoted binding toward DLL4-coated beads, but not JAG1-coated beads. The Notch-ligand assay using Dynabeads suggested that Eogt facilitates DLL4-Notch1 interaction ( Sawaguchi et al., 2017 ).

