| Chapter title |
Monitoring Protein Kinase Expression and Phosphorylation in Cell Lysates with Antibody Microarrays
|
|---|---|
| Chapter number | 9 |
| Book title |
Kinase Screening and Profiling
|
| Published in |
Methods in molecular biology, January 2016
|
| DOI | 10.1007/978-1-4939-3073-9_9 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-3072-2, 978-1-4939-3073-9
|
| Authors |
Hong Zhang, Xiaoqing Shi, Steven Pelech |
| Abstract |
Fuelled by advances in our understanding of the human kinome and phosphoproteome and the increasing availability of pan- and phosphosite-specific antibodies, antibody microarrays have emerged as powerful tools for interrogating protein phosphorylation-mediated signaling systems in ex vivo studies. This economical platform permits ultra-sensitive, semiquantitative measurements of the levels of hundreds of protein kinases and their substrates along with their phosphorylation status simultaneously with minute amounts of specimens. Recent technological innovations in the design and fabrication of antibody microarrays and sample preparation have permitted further refinements of the technology to yield improvements in data quality. In this chapter, we describe a detailed protocol that we have developed for tracking the expression and phosphorylation of protein kinases and their substrates in crude cell lysate samples using a high-content antibody microarray. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 3 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 2 | 67% |
| Researcher | 1 | 33% |
| Readers by discipline | Count | As % |
|---|---|---|
| Pharmacology, Toxicology and Pharmaceutical Science | 1 | 33% |
| Agricultural and Biological Sciences | 1 | 33% |
| Medicine and Dentistry | 1 | 33% |