| Chapter title |
Notch-ligand binding assays in Drosophila cells.
|
|---|---|
| Chapter number | 21 |
| Book title |
Notch Signaling
|
| Published in |
Methods in molecular biology, January 2014
|
| DOI | 10.1007/978-1-4939-1139-4_21 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-1138-7, 978-1-4939-1139-4
|
| Authors |
Aiguo Xu, Kenneth D Irvine, Kenneth D. Irvine, Xu, Aiguo, Irvine, Kenneth D. |
| Abstract |
Activation of the Drosophila transmembrane receptor protein Notch is induced by association with its transmembrane ligands, Delta and Serrate. The ability to assay binding between Notch and its ligands has been essential for characterizing the influence of posttranslational modifications, such as glycosylation, as well as for characterizing structural motifs involved in receptor-ligand interactions. We describe here a simple, widely used method for assaying receptor-ligand binding. This method involves expression of soluble forms of either Notch or its ligands, comprising the extracellular domains fused to an easily assayed tag, the enzyme alkaline phosphatase. These soluble proteins are then incubated with their binding partners, either as transmembrane proteins expressed on the surface of cultured cells or as extracellular protein domains attached to agarose beads. After washing, the amount of bound protein can be readily assayed by measuring alkaline phosphatase activity. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 3 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Professor | 1 | 33% |
| Student > Ph. D. Student | 1 | 33% |
| Student > Doctoral Student | 1 | 33% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 2 | 67% |
| Medicine and Dentistry | 1 | 33% |