Chapter title |
The Peptide Microarray-Based Resonance Light Scattering Assay for Sensitively Detecting Intracellular Kinase Activity.
|
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Chapter number | 7 |
Book title |
Peptide Microarrays
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Published in |
Methods in molecular biology, January 2016
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DOI | 10.1007/978-1-4939-3037-1_7 |
Pubmed ID | |
Book ISBNs |
978-1-4939-3036-4, 978-1-4939-3037-1
|
Authors |
Tao Li, Xia Liu, Dianjun Liu, Zhenxin Wang |
Editors |
Marina Cretich, Marcella Chiari |
Abstract |
The peptide microarray technology is a robust, reliable, and efficient technique for large-scale determination of enzyme activities, and high-throughput profiling of substrate/inhibitor specificities of enzymes. Here, the activities of cyclic adenosine monophosphate (cAMP)-dependent protein kinase A (PKA) in different cell lysates have been detected by a peptide microarray-based resonance light scattering (RLS) assay with gold nanoparticle (GNP) probes. Highly sensitive detection of PKA activity in 0.1 μg total cell proteins of SHG-44 (human glioma cell) cell lysate (corresponding to 200 cells) is achieved by a selected peptide substrate. The experimental results also demonstrate that the RLS assay can be employed to evaluate the chemical regulation of intracellular kinase activity. |
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