Quantitative Western Blot Analysis

Cytosolic protein extracts were isolated from liver tissue and separated in SDS-PAGE as detailed before [34 (link)]. After transferring proteins to a polyvinylidene difluoride membrane (Bio-Rad Laboratories, Hercules, CA, USA), and incubating them with primary antibodies detecting cleaved caspase 3 (Cell Signaling Technology, Massachusetts, USA) or β-actin (Santa Cruz Biotechnology, Dallas, Texas, United States), bands were detected using Super Signal West Dura kit (Thermo Fisher Scientific, Waltham, MA, USA) [34 (link), 36 (link)] and analyzed using the ChemiDoc XRS System with Image Lab software (Bio-Rad Laboratories, Hercules, CA, USA).

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Rajcic D., Brandt A., Jin C.J., Sánchez V., Engstler A.J., Jung F., Nier A., Baumann A, & Bergheim I. (2020). Exchanging dietary fat source with extra virgin olive oil does not prevent progression of diet-induced non-alcoholic fatty liver disease and insulin resistance. PLoS ONE, 15(9), e0237946.

Publication 2020

polyvinylidene difluoride membrane cleaved caspase 3 β-actin Super Signal West Dura kit ChemiDoc XRS System Image Lab software

Actin Antibodies Caspase 3 Cytosolic Dura Liver Membrane proteins Polyvinylidene difluoride Protein Sds page Tissue

Corresponding Organization : Friedrich Schiller University Jena

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Levels of cleaved caspase 3 protein

control variables

Levels of β-actin protein (used as a loading control)

controls

Positive control: Not specified
Negative control: Not specified

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