Western Blot Analysis of Phospho-MAPK Signaling

Protein expression was measured by Western blot as previously described (Siti et al., 2021 (link)). Anti-phospho-ERK1/2 rabbit polyclonal (1:1,000) (#4377), anti-phospho-JNK1/2 rabbit monoclonal (1:1,000) (#4668), and anti-phospho-p38 mouse monoclonal (1:500) (sc-166182; Santa Cruz Biotechnology, Dallas, TX, United States) were the primary antibodies used in this study. β-Actin mouse monoclonal antibodies (1:500) (sc-47778; Santa Cruz Biotechnology, Dallas, TX, United States) served as the loading control. HRP-conjugated IgG anti-mouse (1:2000) (sc-516102; Santa Cruz Biotechnology, Dallas, TX, United States) was used as the secondary antibody. Blots were visualized on a gel doc system and analyzed with ImageJ software (U. S. National Institutes of Health, Bethesda, MD, United States). A minimum of three biological replicates was performed in triplicate (n = 3).

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Siti H.N., Jalil J., Asmadi A.Y, & Kamisah Y. (2021). Parkia speciosa Hassk. Empty Pod Extract Alleviates Angiotensin II-Induced Cardiomyocyte Hypertrophy in H9c2 Cells by Modulating the Ang II/ROS/NO Axis and MAPK Pathway. Frontiers in Pharmacology, 12, 741623.

Publication 2021

sc-166182 β-Actin mouse monoclonal antibodies

Actin Antibodies Antibody Biological Erk1 Monoclonal antibodies Mouse Protein Rabbit Western blot

Corresponding Organization : National University of Malaysia

Other organizations : Sultan Zainal Abidin University, National Defence University of Malaysia

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Variable analysis

independent variables

Stimuli/treatments applied to the cells/samples

dependent variables

Protein expression levels of phospho-ERK1/2, phospho-JNK1/2, and phospho-p38 measured by Western blot

control variables

Loading control: β-Actin
Secondary antibody: HRP-conjugated IgG anti-mouse

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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