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Assays for tyrosine phosphorylation in human cells
- Source :
- Methods Enzymol
- Publication Year :
- 2019
- Publisher :
- Elsevier, 2019.
-
Abstract
- Tyrosine kinases are important for many cellular processes and disruption of their regulation is a factor in diseases like cancer, therefore they are a major target of anticancer drugs. There are many ways to measure tyrosine kinase activity in cells by monitoring endogenous substrate phosphorylation, or by using peptide substrates and incubating them with cell lysates containing active kinases. However, most of these strategies rely on antibodies and/or are limited in how accurately they model the intracellular environment. In cases in which activity needs to be measured in cells, but endogenous substrates are not known and/or suitable phosphospecific antibodies are not available, cell-deliverable peptide substrates can be an alternative and can provide information on activation and inhibition of kinases in intact, live cells. In this chapter, we review this methodology and provide a protocol for measuring Abl kinase activity in human cells using enzyme-linked immunosorbent assay (ELISA) with a generic antiphosphotyrosine antibody for detection.
- Subjects :
- 0303 health sciences
ABL
Kinase
030303 biophysics
Enzyme-Linked Immunosorbent Assay
Tyrosine phosphorylation
Article
Substrate Specificity
Cell biology
03 medical and health sciences
chemistry.chemical_compound
Substrate-level phosphorylation
chemistry
Cell-penetrating peptide
Humans
Tyrosine
Phosphorylation
Kinase activity
K562 Cells
Proto-Oncogene Proteins c-abl
Tyrosine kinase
K562 cells
Subjects
Details
- Database :
- OpenAIRE
- Journal :
- Methods Enzymol
- Accession number :
- edsair.doi.dedup.....e096703c0f83160204347c3ccbea93b7