Hippocampal Protein Extraction and Blotting

Protein extraction from the hippocampal tissue and blotting analyses were performed as previously illustrated.40 (link) Nuclear factor erythroid 2-related factor 2 (Nrf2) (MAB3925, 1:1000; R&D System), heme oxygenase-1 (HO-1) (sc-390991, 1:1000; Santa Cruz Biotechnology, Santa Cruz, CA, USA), and β-actin (MAB8929, 1:2000; R&D System, Minneapolis, MN, USA) were used as a primary antibody, whereas goat anti-mouse IgG (sc-2039, 1:5000; Santa Cruz Biotechnology, Santa Cruz, CA, USA) conjugated with horseradish peroxidase (HRP) was used a secondary antibody. The protein bands were visualized using a chemiluminescence HRP substrate (Bio-Rad, Hercules, CA, USA). The blot intensity was then quantified using Image J software referenced to β-actin.

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Yuan X., Fu Z., Ji P., Guo L., Al-Ghamdy A.O., Alkandiri A., Habotta O.A., Abdel Moneim A.E, & Kassab R.B. (2020). Selenium Nanoparticles Pre-Treatment Reverse Behavioral, Oxidative Damage, Neuronal Loss and Neurochemical Alterations in Pentylenetetrazole-Induced Epileptic Seizures in Mice. International Journal of Nanomedicine, 15, 6339-6353.

Publication 2020

MAB3925 sc-390991 MAB8929 goat anti-mouse IgG (sc-2039, chemiluminescence HRP substrate

Actin Anti igg Antibody Chemiluminescence Goat Heme oxygenase 1 Horseradish peroxidase Mouse Nrf2 Protein Tissue

Corresponding Organization : Shandong First Medical University

Other organizations : First Affiliated Hospital of Zhengzhou University, Union Hospital, Al Baha University, Public Authority for Applied Education and Training, Mansoura University, Helwan University

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Levels of nuclear factor erythroid 2-related factor 2 (Nrf2)
Levels of heme oxygenase-1 (HO-1)

control variables

Protein extraction from the hippocampal tissue
Western blotting analysis procedure
Use of β-actin as a loading control

controls

Positive control: None mentioned
Negative control: None mentioned

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