Chapter title |
Method for Assaying the Lipid Kinase Phosphatidylinositol-5-phosphate 4-kinase α in Quantitative High-Throughput Screening (qHTS) Bioluminescent Format
|
---|---|
Chapter number | 1 |
Book title |
Lipid Signaling Protocols
|
Published in |
Methods in molecular biology, January 2016
|
DOI | 10.1007/978-1-4939-3170-5_1 |
Pubmed ID | |
Book ISBNs |
978-1-4939-3169-9, 978-1-4939-3170-5
|
Authors |
Mindy I. Davis, Atsuo T. Sasaki, Anton Simeonov |
Abstract |
Lipid kinases are important regulators of a variety of cellular processes and their dysregulation causes diseases such as cancer and metabolic diseases. Distinct lipid kinases regulate the seven different phosphorylated forms of phosphatidylinositol (PtdIns). Some lipid kinases utilize long-chain lipid substrates that have limited solubility in aqueous solutions, which can lead to difficulties in developing a robust and miniaturizable biochemical assay. The ability to prepare the lipid substrate and develop assays to identify modulators of lipid kinases is important and is the focus of this methods chapter. Herein, we describe a method to prepare a DMSO-based lipid mixture that enables the 1536-well screening of the lipid kinase phosphatidylinositol-5-phosphate 4-kinase α (PI5P4Kα) utilizing the D-myo-di16-PtIns(5)P substrate in quantitative high-throughput screening (qHTS) format using the ADP-Glo™ technology to couple the production of ADP to a bioluminescent readout. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 8 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Researcher | 2 | 25% |
Student > Ph. D. Student | 1 | 13% |
Student > Bachelor | 1 | 13% |
Other | 1 | 13% |
Unknown | 3 | 38% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 2 | 25% |
Physics and Astronomy | 1 | 13% |
Chemistry | 1 | 13% |
Unknown | 4 | 50% |