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Alpha technology: A powerful tool to detect mouse brain intracellular signaling events
- Source :
- Journal of Neuroscience Methods, Journal of Neuroscience Methods, Elsevier, 2020, 332, pp.108543. ⟨10.1016/j.jneumeth.2019.108543⟩
- Publication Year :
- 2020
- Publisher :
- HAL CCSD, 2020.
-
Abstract
- Background Phosphorylation by protein kinases is a fundamental molecular process involved in the regulation of signaling activities in living organisms. Understanding this complex network of phosphorylation, especially phosphoproteins, is a necessary step for grasping the basis of cellular pathophysiology. Studying brain intracellular signaling is a particularly complex task due to the heterogeneous complex nature of the brain tissue, which consists of many embedded structures. New method Overcoming this degree of complexity requires a technology with a high throughput and economical in the amount of biological material used, so that a large number of signaling pathways may be analyzed in a large number of samples. We have turned to Alpha (Amplified Luminescent Proximity Homogeneous Assay) technology. Comparison with existing method Western blot is certainly the most commonly used method to measure the phosphorylation state of proteins. Even though Western blot is an accurate and reliable method for analyzing modifications of proteins, it is a time-consuming and large amounts of samples are required. Those two parameters are critical when the goal of the research is to comprehend multi-signaling proteic events so as to analyze several targets from small brain areas. Result Here we demonstrate that Alpha technology is particularly suitable for studying brain signaling pathways by allowing rapid, sensitive, reproducible and semi-quantitative detection of phosphoproteins from individual mouse brain tissue homogenates and from cell fractionation and synaptosomal preparations of mouse hippocampus. Conclusion Alpha technology represents a major experimental step forward in unraveling the brain phosphoprotein-related molecular mechanisms involved in brain-related disorders.
- Subjects :
- 0301 basic medicine
Cell signaling
[SDV]Life Sciences [q-bio]
Blotting, Western
Alpha (ethology)
Kinases
Computational biology
Mice
03 medical and health sciences
0302 clinical medicine
Alpha (Amplified Luminescent Proximity Homogeneous Assay) technology
Western blot
medicine
Animals
Phosphorylation
ComputingMilieux_MISCELLANEOUS
medicine.diagnostic_test
Kinase
Chemistry
General Neuroscience
Brain
Phosphoproteins
030104 developmental biology
AlphaScreen / AlphaLISA SureFire Ultra
Cell fractionation
Signal transduction
030217 neurology & neurosurgery
Intracellular
Signal Transduction
Subjects
Details
- Language :
- English
- ISSN :
- 01650270
- Database :
- OpenAIRE
- Journal :
- Journal of Neuroscience Methods, Journal of Neuroscience Methods, Elsevier, 2020, 332, pp.108543. ⟨10.1016/j.jneumeth.2019.108543⟩
- Accession number :
- edsair.doi.dedup.....01f87eb374796cdc4ae54c789fdd7d59