Protein Extraction and Western Blot Analysis

We performed protein extraction and western blot analyses as described previously [29 (link)]. The utilized antibodies included goat antibody to HO-1 (AF3169, 1:500; R&D System), GAPDH (AF5718, 1:500; R&D System), goat anti-mouse IgG (sc-2039, 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 GAPDH, and the data expressed in terms of percent relative to controls.

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Abdelfattah M.S., Elmallah M.I., Ebrahim H.Y., Almeer R.S., Eltanany R.M, & Abdel Moneim A.E. (2019). Prodigiosins from a marine sponge-associated actinomycete attenuate HCl/ethanol-induced gastric lesion via antioxidant and anti-inflammatory mechanisms. PLoS ONE, 14(6), e0216737.

Publication 2019

AF3169 AF5718 goat anti-mouse IgG (sc-2039 enhanced chemiluminescence detection kit Kodak Image Station 2000R

Anti igg Antibodies Antibody Chemiluminescence Gapdh Goat Mouse Protein Western blot analyses

Corresponding Organization : Helwan University

Other organizations : King Saud University

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Variable analysis

independent variables

Protein extraction and western blot analyses

dependent variables

Protein band intensity

control variables

GAPDH (used for normalizing protein band intensity)

controls

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

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