SDS-PAGE and Immunoblotting of Cultured Cells and Mouse Brains

Cultured cells were processed and analysed by SDS‐PAGE and immunoblotting as described previously (Gomez‐Suaga et al., 2017 (link)). After probing with primary antibodies, the blots were incubated with horseradish peroxidase conjugated secondary antibodies and developed using chemiluminescence with a Luminata Forte Western HRP substrate system according to the manufacturer's instructions (Millipore). Signals were detected using a BioRad ChemiDoc MP Imaging system. Mouse brains were prepared for SDS‐PAGE as described previously (Stoica et al., 2016 (link)), and samples were analysed as above for cultured cells.

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Gomez‐Suaga P., Mórotz G.M., Markovinovic A., Martín‐Guerrero S.M., Preza E., Arias N., Mayl K., Aabdien A., Gesheva V., Nishimura A., Annibali A., Lee Y., Mitchell J.C., Wray S., Shaw C., Noble W, & Miller C.C. (2022). Disruption of ER‐mitochondria tethering and signalling in C9orf72‐associated amyotrophic lateral sclerosis and frontotemporal dementia. Aging Cell, 21(2), e13549.

Publication 2022

Luminata Forte Western HRP substrate system ChemiDoc MP Imaging system

Antibodies Brains Chemiluminescence Cultured cells Mouse Sds page

Corresponding Organization : King's College London

Other organizations : University College London, National Hospital for Neurology and Neurosurgery, UK Dementia Research Institute

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Variable analysis

independent variables

Cultured cells processed and analyzed by SDS-PAGE and immunoblotting
Mouse brains prepared for SDS-PAGE and analyzed

dependent variables

Signals detected using a BioRad ChemiDoc MP Imaging system

control variables

Experimental procedures described previously in Gomez-Suaga et al. (2017) and Stoica et al. (2016)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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