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Western Blot Analysis

JH Jun He
YH Ying He
BY Bing Yu
XW Xuelian Wang
DC Daiwen Chen
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Protein extracts from muscle samples were separated by 10% SDS-PAGE after adding 5 × Laemmli sample buffer and boiling. The separated proteins were transferred onto a PVDF membrane and probed with rabbit polyclonal antibodies against slow-MHC (ab11083, Abcam), fast-MHC (ab91506, Abcam), MYOD1 (ab16148, Abcam), MYOG (ab1835, Abcam), and p21 (ab109199; Abcam). Horseradish peroxidase (HPRP)-conjugated goat anti-rabbit IgG (Santa Cruz Biotechnology) was used as the secondary antibody. For quantification, glyceraldehyde 3-phosphate dehydrogenase (ab8245, Abcam) or tubulin (ab52866, Abcam) was selected as the internal standard. Bound antibodies were detected using the ECL Prime western blotting detection reagent (GE Healthcare) on a CCD-based imager (ImageQuant LAS 4000, GE Healthcare).

Copyright and License information:  ©2020 He, He, Yu, Wang and Chen.
This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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