Western Blot Assay for AMPK Signaling

Western blot assay was carried out referring to a previous publication [24 (link)]. Antibodies, including AMPKα (cat. no.2532), phosphor-AMPKα (Thr172) (cat. no. 2531), ACC (cat. no.3662), phosphor-ACC (Ser79) (cat. no.3661), and O-GlcNAc (cat. no.9875), were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibodies (cat. no. 309154) and HRP-conjugated secondary antibodies were purchased from Golden Bridge Biotechnology (Beijing, China).

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Pang Y., Xu X., Xiang X., Li Y., Zhao Z., Li J., Gao S., Liu Q., Mai K, & Ai Q. (2021). High Fat Activates O-GlcNAcylation and Affects AMPK/ACC Pathway to Regulate Lipid Metabolism. Nutrients, 13(6), 1740.

Publication 2021

O-GlcNAc

Anti antibodies Antibodies Glyceraldehyde 3 phosphate dehydrogenase Phosphor Western blot assay

Corresponding Organization : Qingdao National Laboratory for Marine Science and Technology

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Protein expression levels of AMPKα, phosphor-AMPKα (Thr172), ACC, phosphor-ACC (Ser79), and O-GlcNAc

control variables

Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as a loading control

controls

Positive control: None mentioned
Negative control: None mentioned

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