pampka (thr172, 2535) antibody  (Cell Signaling Technology Inc)

Journal: Molecular metabolism

Article Title: Mouse tissue harvest-induced hypoxia rapidly alters the in vivo metabolome, between-genotype metabolite level differences, and 13 C-tracing enrichments.

doi: 10.1016/j.molmet.2022.101596

Figure Lengend Snippet: Figure 2: The post-dissection liver metabolome is rapidly remodeled by hypoxia. (A) Schematic depicting the dependence of the NADH oxidation and O2 reduction activities of the electron transport chain (ETC) on the ATP synthesis activity of the ATP synthase/Complex V (V). (BeC) Line graphs showing the NADþ and NADH (B) and AMP, ADP, and ATP (C) metabolite fold changes by delayed freezing time-point compared to time zero. Data are presented as the mean SEM. P-values were calculated by one-way ANOVA with the post-hoc Holm-Sidak multiple comparison test (n ¼ 6 biological replicates). (D) Representative Western blot of liver tissue p-AMPK (Thr172), AMPK, and Hsp90 by delayed freezing time-point and a line graph showing p-AMPK/AMPK quantification. Quantified densiometric data are presented as the mean SEM. P-values were calculated by one-way ANOVA with the post-hoc Holm-Sidak multiple comparison test (n ¼ 6 biological replicates). *p < 0.05, **p < 0.01, ***p < 0.001; Related to Figure S2 and Table S1.

Article Snippet: Blocked membranes were incubated with primary antibodies at 4 C overnight [mouse monoclonal anti-AMPKa (F6) (RRID: AB_915794), rabbit monoclonal anti-pAMPKa (Thr172) (40H9) (RRID: AB_331250), and rabbit monoclonal anti-HSP90 (RRID: AB_2121214) from Cell Signaling Technology].

Techniques: Dissection, Activity Assay, Comparison, Western Blot