Phos-tag-based non-radioactive protocols for monitoring Arabidopsis kinase activities in vitro
Summary: Kinases are indispensable signaling components. Radioactive-based phosphorylation assays are widely used but require specific protective equipment and safety trainings. Here, we present a Phos-tag-based non-radioactive kinase assay to study Arabidopsis kinase activities. We expressed and pu...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2022-12-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166722005974 |
| Summary: | Summary: Kinases are indispensable signaling components. Radioactive-based phosphorylation assays are widely used but require specific protective equipment and safety trainings. Here, we present a Phos-tag-based non-radioactive kinase assay to study Arabidopsis kinase activities. We expressed and purified both kinase and substrate proteins from E. coli cells and then used the Phos-tag technology to detect the kinase activities under either different temperatures or chemical treatments. This non-radioactive approach is environmentally friendly and applicable to other kinases and organisms.For complete details on the use and execution of this protocol, please refer to Lin et al. (2022). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
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| ISSN: | 2666-1667 |