Protocol detail

Western Blot Analysis Protocol

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Western blot analyses were performed as described previously^{33 (link)}. Briefly, cells were lysed using RIPA buffer (Invitrogen) containing SIGMAFAST Protease Inhibitor Cocktail (Sigma) and PhoSTOP phosphatase inhibitors (Roche). Adipose tissue lysates were prepared using PRO-PREP Protein Extraction Solution (iNtRON Biotechnology). Primary antibodies used for western blot analyses were described in Supplementary Tables S3. NIH ImageJ software was used for the quantification. Full-length immunoblot membrane images are attached in Supplementary Information.

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Son Y., Choi C., Song C., Im H., Cho Y.K., Son J.S., Joo S., Joh Y., Lee Y.J., Seong J.K, & Lee Y.H. (2021). Development of CIDEA reporter mouse model and its application for screening thermogenic drugs. Scientific Reports, 11, 18429.

Publication 2021

RIPA buffer SIGMAFAST Protease Inhibitor Cocktail PhoSTOP phosphatase inhibitors PRO-PREP Protein Extraction Solution

Adipose tissue Antibodies Buffer Cells Immunoblot Inhibitors Intron Membrane Phosphatase Pro protein Protease inhibitor Ripa Western blot analyses

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Variable analysis

independent variables

Manipulation of cell lysis using RIPA buffer and protease/phosphatase inhibitors

dependent variables

Protein expression levels measured by Western blot analyses

control variables

Primary antibodies used for Western blot analyses (described in Supplementary Tables S3)
Quantification method using NIH ImageJ software

controls

Full-length immunoblot membrane images provided as Supplementary Information (presumably for positive and negative controls)

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