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Children's Hospital of Eastern Ontario Research Institute

3 ARTICLES PUBLISHED IN JoVE

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Biology

Identification of Kinase-substrate Pairs Using High Throughput Screening
Courtney Reeks 1, Robert A. Screaton 2,3
1Children's Hospital of Eastern Ontario Research Institute, 2Sunnybrook Research Institute, University of Toronto, 3Department of Biochemistry, University of Toronto

Protein phosphorylation is a central feature of how cells interpret and respond to information in their extracellular milieu. Here, we present a high throughput screening protocol using kinases purified from mammalian cells to rapidly identify kinases that phosphorylate a substrate(s) of interest.

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Biology

Isolation of CD146+ Resident Lung Mesenchymal Stromal Cells from Rat Lungs
Jennifer J.P. Collins 1,2, Marius A. Möbius 1,3,4, Bernard Thébaud 1,2,5
1Sinclair Centre for Regenerative Medicine, Ottawa Hospital Research Institute, 2University of Ottawa, 3Department of Neonatology and Pediatric Critical Care Medicine, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, 4DFG Research Center and Cluster of Excellence for Regenerative Therapies (CRTD), Technische Universität, Dresden, 5Children's Hospital of Eastern Ontario Research Institute

This protocol describes an isolation technique for obtaining primary lung resident mesenchymal stromal cells from rats, through the use of enzymatic digestion, density gradient separation, plastic adherence and CD146+ magnetic bead selection.

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Genetics

High-throughput DNA Extraction and Genotyping of 3dpf Zebrafish Larvae by Fin Clipping
Ceres Kosuta 1,2, Kate Daniel 1, Devon L. Johnstone 1,2, Kevin Mongeon 1,2, Kevin Ban 1,2, Sophie LeBlanc 1, Stuart MacLeod 1, Karim Et-Tahiry 2, Marc Ekker 2, Alex MacKenzie 1, Izabella Pena 1,2
1Children's Hospital of Eastern Ontario Research Institute, 2Department of Biology, University of Ottawa

Zebrafish have been used as reliable genetic model organisms in biomedical research, especially with the advent of gene-editing technologies. When larval phenotypes are expected, DNA extraction and genotype identification can be challenging. Here, we describe an efficient genotyping procedure for zebrafish larvae, by tail clipping, as early as 72-h post-fertilization.

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