Thioacetamide (TAA) was purchased from Sigma-Aldrich (St. Louis, MO, USA) and dissolved in normal saline (0.9% sodium chloride). Primary antibodies against α-SMA, collagen-I, Smad4, and B-cell lymphoma 2 (Bcl-2)-associated X protein (BAX) were purchased from Abcam (Cambridge, UK). ER-α, Smad7, Bcl-2, p53, and β-actin were obtained from Santa Cruz Biotechnology (Dallas, TX, USA). Vimentin, p-Smad2/3, Smad2/3, cleaved caspase 3, p-PI3K, p-Akt, phosphatase and tensin homolog (PTEN), and p-PTEN were purchased from Cell Signaling Technology (Danvers, MA, USA). Assay kits used to measure aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), gamma-glutamyl transferase (γ-GGT), malondialdehyde (MDA), and catalase (CAT) were purchased from Abcam (Cambridge, UK). Total glutathione (GSH) assay kits were purchased from Enzo Life Sciences (New York, NY, USA). SOD assay kits were purchased from Cayman Chemical (Ann Arbor, MI, USA). Hydroxyproline assay kits were purchased from Cell Biolabs, Inc. (San Diego, CA, USA). All other chemicals used in this study were purchased from Sigma-Aldrich (St. Louis, MO, USA).
Catalase cat
Manufactured by Abcam
Sourced in United Kingdom, United States
Catalase (CAT) is an enzyme that catalyzes the decomposition of hydrogen peroxide (H2O2) into water and oxygen. It is a crucial antioxidant enzyme found in nearly all living organisms that are exposed to oxygen, where it functions to protect cells from oxidative damage by reactive oxygen species.
Automatically generated - may contain errors
Lab products found in correlation
Smad7, by Santa Cruz Biotechnology (1 mentions)
Thioacetamide taa, by Merck Group (1 mentions)
Primary antibodies against α sma, by Abcam (1 mentions)
Smad2 3, by Cell Signaling Technology (1 mentions)
P pten, by Cell Signaling Technology (1 mentions)
Sod assay kit, by Cayman Chemical (1 mentions)
Phosphatase and tensin homolog pten, by Cell Signaling Technology (1 mentions)
P smad2 3, by Cell Signaling Technology (1 mentions)
P akt, by Cell Signaling Technology (1 mentions)
Cleaved caspase 3, by Cell Signaling Technology (1 mentions)
Vimentin, by Cell Signaling Technology (1 mentions)
P pi3k, by Cell Signaling Technology (1 mentions)
Collagen 1, by Abcam (1 mentions)
β actin, by Santa Cruz Biotechnology (1 mentions)
Bcl 2, by Santa Cruz Biotechnology (1 mentions)
Malondialdehyde mda, by Abcam (1 mentions)
Gs 800 imaging systems scanner, by Bio-Rad (1 mentions)
Quantity one 4, by Bio-Rad (1 mentions)
Ecl system, by Bio-Rad (1 mentions)
3 protocols using catalase cat
1
Thioacetamide-Induced Liver Fibrosis Model
2
Oxidative Stress Biomarker Assays
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Malondialdehyde (MDA, cat no. ab118970), Superoxide Dismutase (SOD, ab65354), Catalase (CAT, cat no. ab83464), Glutathione reductase (GRx, cat no. ab239727), Glutathione Peroxidase (GPx, cat no. ab102530), and Hydrogen Peroxide (H2O2, cat no. ab102500) assay kits were obtained from Abcam Inc. Waltham, MA, USA.
3
Validating Quantitative Proteomic Findings via Immunoblotting
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Immunoblotting was performed to validate the findings of the mass spectrometry-based quantitative proteomic analysis. The proteins extracted from OC and CTR cells were mixed in six pools (3 OC and 3 CTR, same quantity of each sample), separated by SDS-PAGE 10% or 12%, depending on the molecular mass of protein under evaluation) and blotted on PDVF membranes. The blotted membranes were incubated with antibodies against actin (ACT) (Sigma-Aldrich, 1:5000), betahexosaminidase subunit beta (HEXB) (Santa Cruz, 1:1000), mitochondrial superoxide dismutase (SOD2), (Cell Signaling, 1:5000), collagenase 3 (MMP13) (Santa Cruz, 1:1000), cytochrome P450 1B1 (CYP1B1) (Santa Cruz, 1:1000), catalase (CAT) (Abcam, 1:2000) and tubulin (TUB) (Santa Cruz, 1:5000) at 4 °C overnight and then with the appropriate anti-mouse HRP secondary antibody (Sigma-Aldrich, 1:5000) or anti-rabbit HRP secondary antibody (Sigma-Aldrich 1:5000). Immunoreactivity was detected by chemiluminescence using the ECL system (Bio-Rad). Images were acquired using a GS-800 imaging systems scanner (Bio-Rad). A densitometric analysis was performed with Quantity One 4.5.0 software (Bio-Rad) using tubulin bands as normalization factor. The t-test was carried out for the statistical analysis where p ≤ 0.05 was considered significant.
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