Nrf2 and HO-1 Protein Analysis

Protein extraction and blotting analyses were carried out as previously reported [35 (link)]. The utilized antibodies included mouse antibody to Nrf2 (MAB3925, 1:500; R&D System), HO-1 (sc-390991, 1:750; Santa Cruz Biotechnology, Santa Cruz, CA, USA), β-actin (MAB8929, 1:500; R&D System), and goat anti-mouse IgG (sc-2039, 1:5000; Santa Cruz Biotechnology, Santa Cruz, CA, USA). The proteins were visualized using an enhanced chemiluminescence detection kit (Bio-Rad, Hercules, CA, USA) following the manufacturer’s protocol. Images were analyzed using the Kodak Image Station 2000R (Eastman Kodak Company, Rochester, NY, USA). Protein bands intensity were referenced to β-actin, and the data presented in terms of percent relative to controls.

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AL-Megrin W.A., Alkhuriji A.F., Yousef A.O., Metwally D.M., Habotta O.A., Kassab R.B., Abdel Moneim A.E, & El-Khadragy M.F. (2019). Antagonistic Efficacy of Luteolin against Lead Acetate Exposure-Associated with Hepatotoxicity is Mediated via Antioxidant, Anti-Inflammatory, and Anti-Apoptotic Activities. Antioxidants, 9(1), 10.

Publication 2019

MAB3925 sc-390991 MAB8929 sc-2039 enhanced chemiluminescence detection kit Kodak Image Station 2000R

Actin Anti igg Antibodies Antibody Chemiluminescence Goat Mouse Nrf2 Protein

Corresponding Organization : Helwan University

Other organizations : King Saud University, Zagazig University, Mansoura University

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Protein expression levels of Nrf2, HO-1, and β-actin

control variables

None explicitly mentioned

controls

Positive control: β-actin, used to normalize protein expression levels
Negative control: Not specified

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