HUVECs or thoracic aorta were lysed in lysis buffer (Merck Millipore) with protease inhibitor cocktail (Roche) and phosphatase inhibitor cocktail (Roche). Protein concentrations were quantified using a BCA Kit (Beyotime). Portions of lysate with equal amount of protein were subjected to SDS-PAGE and then electrotransferred to a polyvinylidene fluoride membrane (Merck Millipore). After being blocked with 10% bovine serum albumin (BSA), the membrane was incubated with primary antibodies (1:1000 or 1:500) overnight at 4 °C, followed by incubation with peroxidase-conjugated goat anti-rabbit or anti-mouse IgG (1:5000) for 1 h at room temperature, and then detected by enhanced chemiluminescence (Thermo Scientific) and imaged using a ChemiScope Mini chemiluminescence imaging system (Clinx Science). Band intensities were quantified using the software ImageJ (National Institutes of Health).
Chemiscope mini chemiluminescence imaging system
Manufactured by Clinx
The ChemiScope Mini is a chemiluminescence imaging system designed for versatile and compact visualization of biological samples. The system captures high-quality images of chemiluminescent signals from a variety of applications, including Western blots, EMSA, and reporter gene assays.
Automatically generated - may contain errors
Lab products found in correlation
Polyvinylidene fluoride membrane, by Merck Group (2 mentions)
Bca kit, by Beyotime (2 mentions)
Lysis buffer, by Merck Group (2 mentions)
Protease inhibitor cocktail, by Roche (2 mentions)
Phosphatase inhibitor cocktail, by Roche (1 mentions)
Enhanced chemiluminescence, by Thermo Fisher Scientific (1 mentions)
Wbkls0500, by Merck Group (1 mentions)
F1804, by Merck Group (1 mentions)
T6074, by Merck Group (1 mentions)
G9295, by Merck Group (1 mentions)
Ab62352, by Abcam (1 mentions)
2 protocols using chemiscope mini chemiluminescence imaging system
1
Western Blot Analysis of Protein Expression
2
Western Blotting Comprehensive Protocol
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Cells were lysed using lysis buffer (Millipore) with Protease Inhibitor Cocktail (Roche). Protein quantification was performed using a BCA Kit (Beyotime). Protein lysate was subjected to SDS‐PAGE and subsequently electrotransferred onto a polyvinylidene fluoride membrane (Millipore). Blots were developed by indicated antibodies and enhanced chemiluminescence (ECL) (Millipore, WBKLS0500), followed by a ChemiScope Mini chemiluminescence imaging system (Clinx Science). The antibodies used are listed as follows: anti‐GPx7 (Abclone, A3902, 1:1,000), anti‐Nrf2 (Abcam, ab62352, 1:1,000), anti‐PDI (Abcam, ab2792, 1:2,000), anti‐GPx8 (GeneTex, GTX125992, 1:1,000), anti‐Prx4 (Animal Facility, Institute of Genetics and Developmental Biology, CAS, rabbit serum, 1:500), anti‐ERp44 (CST, 3798, 1:2,000), anti‐Ero1α (Millipore, MABT376, 1:1,000), anti‐ERp46 (Animal Facility, Institute of Genetics and Developmental Biology, CAS, rabbit serum, 1:500), anti‐ERp72 (Origene, TA503904, 1:2,000), anti‐GAPDH (Sigma, G9295, 1:50,000), anti‐Flag (Sigma, F1804, 1:4,000), anti‐α‐tubulin (Sigma, T6074, 1:10,000), goat anti‐rabbit IgG (Sigma, A0545, 1:10,000), goat anti‐mouse IgG (Sigma, A4416, 1:10,000).
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