| Chapter title |
Bioluminescence Methods for Assaying Kinases in Quantitative High-Throughput Screening (qHTS) Format Applied to Yes1 Tyrosine Kinase, Glucokinase, and PI5P4Kα Lipid Kinase
|
|---|---|
| Chapter number | 4 |
| Book title |
Kinase Screening and Profiling
|
| Published in |
Methods in molecular biology, January 2016
|
| DOI | 10.1007/978-1-4939-3073-9_4 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-3072-2, 978-1-4939-3073-9
|
| Authors |
Mindy I. Davis, Douglas S. Auld, James Inglese |
| Abstract |
Assays in which the detection of a biological phenomenon is coupled to the production of bioluminescence by luciferase have gained widespread use. As firefly luciferases (FLuc) and kinases share a common substrate (ATP), coupling of a kinase to FLuc allows for the amount of ATP remaining following a kinase reaction to be assessed by quantitating the amount of luminescence produced. Alternatively, the amount of ADP produced by the kinase reaction can be coupled to FLuc through a two-step process. This chapter describes the bioluminescent assays that were developed for three classes of kinases (lipid, protein, and metabolic kinases) and miniaturized to 1536-well format, enabling their use for quantitative high-throughput (qHTS) of small-molecule libraries. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 12 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Unspecified | 2 | 17% |
| Researcher | 2 | 17% |
| Professor > Associate Professor | 2 | 17% |
| Student > Master | 1 | 8% |
| Student > Ph. D. Student | 1 | 8% |
| Other | 1 | 8% |
| Unknown | 3 | 25% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 3 | 25% |
| Unspecified | 2 | 17% |
| Arts and Humanities | 1 | 8% |
| Computer Science | 1 | 8% |
| Medicine and Dentistry | 1 | 8% |
| Other | 0 | 0% |
| Unknown | 4 | 33% |