Western Blot Analysis of Antioxidant Proteins

We performed protein extraction and western blot analyses as described previously^{20 (link)}. The utilized antibodies included mouse antibody to NQO1 (sc-376023, 1:500; Santa Cruz Biotechnology, Santa Cruz, CA, USA), Nrf2 (MAB3925, 1:500; R&D System), NFκB (p65) (sc-8008, 1:200; Santa Cruz Biotechnology), goat antibody to HO-1 (AF3169, 1:500; R&D System), β-actin (MAB8929, 1:500; R&D System), goat anti-mouse IgG (sc-2039, 1:5,000; Santa Cruz Biotechnology, Santa Cruz, CA, USA) and donkey anti-goat IgG (sc-2042, 1:5,000; Santa Cruz Biotechnology, Santa Cruz, CA, USA). The proteins were visualized using an enhanced chemiluminescence detection kit (Bio-Rad, 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 normalized to β-actin, and the data expressed in terms of percent relative to controls.

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Almeer R.S., AlBasher G.I., Alarifi S., Alkahtani S., Ali D, & Abdel Moneim A.E. (2019). Royal jelly attenuates cadmium-induced nephrotoxicity in male mice. Scientific Reports, 9, 5825.

Publication 2019

sc-376023 MAB3925 sc-8008 AF3169 MAB8929 goat anti-mouse IgG sc-2039 donkey anti-goat IgG sc-2042 enhanced chemiluminescence detection kit Kodak Image Station 2000R

Actin Anti igg Antibodies Antibody Chemiluminescence Donkey Goat Mouse Nfκb p65 Nqo1 Nrf2 Protein Western blot analyses

Corresponding Organization : King Saud University

Other organizations : Helwan University

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Variable analysis

independent variables

Protein extraction and western blot analyses methods as described previously in reference 20

dependent variables

Protein expression levels of NQO1, Nrf2, NFκB (p65), and HO-1 measured by western blot

control variables

β-actin protein expression level used for normalization of protein band intensities

controls

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

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