S'identifier

Jewish General Hospital

4 ARTICLES PUBLISHED IN JoVE

Biology

Dithranol as a Matrix for Matrix Assisted Laser Desorption/Ionization Imaging on a Fourier Transform Ion Cyclotron Resonance Mass Spectrometer

Cuong H. Le^1,2, Jun Han¹, Christoph H. Borchers^1,2

¹University of Victoria-Genome BC Proteomics Centre, University of Victoria, ²Department of Biochemistry and Microbiology, University of Victoria

Dithranol (DT; 1,8-dihydroxy-9,10-dihydroanthracen-9-one) has previously been reported as a MALDI matrix for tissue imaging of small molecules; protocols for the use of DT for the MALDI imaging of endogenous lipids on the surface of tissue sections by positive-ion MALDI-MS on an ultrahigh-resolution quadrupole-FTICR instrument are provided here.

Biology

Electrophoretic Mobility Shift Assay (EMSA) for the Study of RNA-Protein Interactions: The IRE/IRP Example

Carine Fillebeen^1,2, Nicole Wilkinson^1,2, Kostas Pantopoulos^1,2

¹Lady Davis Institute for Medical Research, Jewish General Hospital, ²Department of Medicine, McGill University

Here we present a protocol to analyze RNA/protein interactions. The electrophoretic mobility shift assay (EMSA) is based on the differential migration of RNA/protein complexes and free RNA during native gel electrophoresis. By using a radiolabeled RNA probe, RNA/protein complexes can be visualized by autoradiography.

Neuroscience

Modeling Neuronal Death and Degeneration in Mouse Primary Cerebellar Granule Neurons

Matthew Laaper^1,2, Takrima Haque¹, Ruth S. Slack³, Arezu Jahani-Asl^1,2,4

¹Lady Davis Institute for Medical Research, Jewish General Hospital, ²Integrated Program in Neuroscience, McGill University, ³Department of Cellular and Molecular Medicine, University of Ottawa, ⁴Department of Oncology, Faculty of Medicine, McGill University

This protocol describes a simple method for isolating and culturing primary mouse cerebral granule neurons (CGNs) from 6-7 day old pups, efficient transduction of CGNs for loss and gain of function studies, and modelling NMDA-induced neuronal excitotoxicity, low-potassium-induced cell death, DNA-damage, and oxidative stress using the same culture model.

Chemistry

Peptide and Protein Quantification Using Automated Immuno-MALDI (iMALDI)

Huiyan Li¹, Robert Popp¹, Bjorn Frohlich¹, Michael X. Chen², Christoph H. Borchers^1,3,4,5

¹University of Victoria-Genome BC Proteomics Centre, ²Jewish General Hospital, McGill University, ³Dept of Biochemistry and Microbiology, University of Victoria, ⁴Proteomics Centre, Segal Cancer Centre, Lady Davis Institute, Jewish General Hospital, McGill University, ⁵Gerald Bronfman Department of Oncology, Jewish General Hospital

A protocol for the protein quantification in complex biological fluids using automated immuno-MALDI (iMALDI) technology is presented.