Western Blotting Analysis of RBM20

Western blotting analysis was performed as described previously.^{29 (link),30 (link)} Briefly, protein lysed from cells and tissues was separated by SDS-PAGE gel and transferred onto a PVDF membrane. The membrane was probed with primary antibodies against RBM20 (1:1500, rabbit, homemade^{3 (link)}), phosphorylated RBM20 S637 (1:1000, rabbit; Abmart Inc. Shanghai, China), GAPDH (1:1500, rabbit; Cell Signaling Technology, Danvers, Massachusetts, USA; Cat#2118S) served as the protein-loading control. Then, the membrane was probed with horseradish peroxidase-coupled secondary antibodies (1:3000, anti-rabbit; Promega; Cat#W4011) for 1 h. Chemiluminescence images were taken by ChemiDoc system (Bio-Rad, Hercules, CA).

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Zhang Y., Wang C., Sun M., Jin Y., Braz C.U., Khatib H., Hacker T.A., Liss M., Gotthardt M., Granzier H., Ge Y, & Guo W. (2022). RBM20 phosphorylation and its role in nucleocytoplasmic transport and cardiac pathogenesis. The FASEB Journal, 36(5), e22302.

Publication 2022

phosphorylated RBM20 S637 GAPDH horseradish peroxidase-coupled secondary antibodies ChemiDoc system

Antibodies Cells Chemiluminescence Gapdh Horseradish peroxidase Membrane Promega Protein Pvdf Rabbit Sds page Tissues Western blotting analysis

Corresponding Organization : University of Wyoming

Other organizations : Max Delbrück Center, Charité - Universitätsmedizin Berlin, University of Arizona

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Variable analysis

independent variables

Western blotting analysis

dependent variables

Protein expression level of RBM20
Phosphorylation level of RBM20 at serine 637

control variables

GAPDH expression level (used as protein-loading control)

controls

Positive control: Not specified
Negative control: Not specified

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