Cells were placed in triplicate into six-well plates and treated with either PBS or PprI (4 μg/mL) 6 h before 4 Gy irradiation. Tweenty-four hours later, SOD activity, CAT activity and MDA concentrations were determined using a T-SOD assay kit, CAT assay kit, and MDA assay kit, respectively (Nanjing Jiancheng Bioengineering Institute, Nanjing, China).
Cat assay kit
Manufactured by Nanjing Jiancheng
Sourced in China
The CAT assay kit is a laboratory tool used to measure the activity of the enzyme catalase. Catalase is an important antioxidant enzyme found in living organisms that helps break down hydrogen peroxide into water and oxygen. The CAT assay kit provides a quantitative method to determine the catalase activity in biological samples.
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Lab products found in correlation
Sod assay kit, by Nanjing Jiancheng (33 mentions)
Mda assay kit, by Nanjing Jiancheng (27 mentions)
Gsh px assay kit, by Nanjing Jiancheng (13 mentions)
Gsh assay kit, by Nanjing Jiancheng (11 mentions)
Pod assay kit, by Nanjing Jiancheng (11 mentions)
H2o2 assay kit, by Nanjing Jiancheng (6 mentions)
T sod assay kit, by Nanjing Jiancheng (5 mentions)
Apx assay kit, by Nanjing Jiancheng (4 mentions)
Gpx assay kit, by Nanjing Jiancheng (4 mentions)
T aoc assay kit, by Nanjing Jiancheng (3 mentions)
Total sod assay kit, by Nanjing Jiancheng (3 mentions)
Ros assay kit, by Nanjing Jiancheng (3 mentions)
Atp assay kit, by Nanjing Jiancheng (2 mentions)
Plant pod assay kit, by Nanjing Jiancheng (2 mentions)
Total protein quantitative assay kit, by Nanjing Jiancheng (2 mentions)
Microscale mda assay kit, by Nanjing Jiancheng (2 mentions)
Proline assay kit, by Nanjing Jiancheng (2 mentions)
Gsh px, by Nanjing Jiancheng (2 mentions)
Atpase assay kit, by Nanjing Jiancheng (2 mentions)
Superoxide dismutase sod assay kit, by Nanjing Jiancheng (2 mentions)
69 protocols using cat assay kit
1
Radiation-Induced Oxidative Stress Assay
2
Oxidative Stress and Apoptosis Assays
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EGF was purchased from Peprotech (Rocky Hill, NJ, USA). LPS was purchased from Sigma-Aldrich (Saint Louis, MO, USA). Fetal bovine serum (FBS), Trypsin/EDTA and antibiotics (Penicillin-Streptomycin for Cell Culture) were from GIBCO (Carlsbad, CA, USA). Dulbecco’s modified Eagle’s F12 Ham medium (HyCloneTM DMEM/F12 1:1 media) was purchased from GE Healthcare life sciences (South Logan, UT, USA). Plastic culture plates were manufactured by Corning Inc. (Corning, NY, USA). CCK-8 Assay Kit, BCA protein assay reagent, LDH Assay Kit, T-AOC Assay Kit, CAT Assay Kit, GSH-Px Assay Kit, SOD Assay Kit, and MDA Assay Kit were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). PBS, RIPA Lysis Buffer R2220 were purchased from Solarbio (Beijing, China). TRIzol Reagent was obtained from Invitrogen (Carlsbad, CA, USA). Annexin V-FITC/PI kits was obtained from Keygen Biotech (Nanjing, China).The primary antibodies against Nrf2, HO-1, NQO1, P53, Bax, Bcl2, Caspase3, β-actin, and the secondary antibody Goat Anti-Rabbit IgG/HRP used in Western blot analyses were all purchased from Proteintech (Rosemont, IL, USA). The primary antibody against Fas was purchased from Abcam (Cambridge, MA, USA).
3
Antioxidant Enzyme Activity Assay
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The activity of antioxidase SOD, GSH-px, and CAT was evaluated by biochemical assays. MC3T3-E1 cells were washed three times with PBS and then lysed with 150 μL RIPA lysate on ice for 1 h, before being centrifuged at 4 °C, 12,000 g for 10 min. In accordance with the instructions of the three enzymes SOD, GSH-px, and CAT assay kit (Nanjing Jiancheng, A001–3, A005, A007-1), the supernatant was removed for detection.
4
Enzymatic Assays for Oxidative Stress
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Superoxide dismutase (SOD), catalase (CAT) and glutathione (GSH) activities in serum samples or mouse livers were analyzed using a SOD assay kit, a CAT assay kit and a GSH assay kit (Jiancheng Bioengineering Institute), respectively. All experiments were carried out according to the manufacturer’s instructions.
5
Antioxidant Enzyme Activity Assay
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SOD assay kit (A001‐3‐2), catalase (CAT) assay kit (A007‐1‐1), and glutathione (GSH) assay kit (A006‐2‐1) were ordered from Nanjing Jiancheng Bioengineering Institute. The serum samples and cell lysates were collected and reacted with corresponding reagents at 37°C (20 min for SOD detection), and the optical density of each well at 450 nm (for SOD) and 405 nm (for CAT) was measured by a microplate reader to assess the activities of SOD and CAT.
To test the level of GSH, the Reagents 1, 2, 3, and 4 were prepared using the GSH assay kit. Fifty microliters serum samples and 100 μL cell lysates were treated with Reagent 1, followed by centrifugation. Then, 100 μL supernatant was collected, and cultured with Reagent 2 (100 μL) and Reagent 3 (25 μL) for 5 min at room temperature. The absorbance of each well at 405 nm was measured by a microplate reader.
To test the level of GSH, the Reagents 1, 2, 3, and 4 were prepared using the GSH assay kit. Fifty microliters serum samples and 100 μL cell lysates were treated with Reagent 1, followed by centrifugation. Then, 100 μL supernatant was collected, and cultured with Reagent 2 (100 μL) and Reagent 3 (25 μL) for 5 min at room temperature. The absorbance of each well at 405 nm was measured by a microplate reader.
6
Oxidative Stress Markers in Hippocampus
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Malondialdehyde (MDA) level in hippocampal tissue was measured by MDA assay kit (TBA method) (Nanjing Jiancheng, Nanjing, China). Glutathione (GSH) level was measured by reduced GSH assay kit (Nanjing Jiancheng). The activity of total superoxide dismutase (T-SOD) was evaluated by T-SOD assay kit (Hydroxylamine method) (Nanjing Jiancheng). The activity of catalase (CAT) was evaluated by CAT assay kit (Nanjing Jiancheng). All the kits were used based on the manufacturer’s instructions.
7
Liver Oxidative Stress Biomarkers Analysis
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Liver tissue samples were homogenized in nine volumes of ice-cold 50 mM phosphate buffer (pH 7.4) and centrifuged at 3,200 × g for 20 min at 4°C. Supernatants were used to determine SOD, GSH-Px, MDA, CAT and total protein concentrations by using commercially available diagnostic kits (SOD assay kit; cat no. A001-3; GSH-PX assay kit; cat no. A005; cat no. MDA assay kit; cat no. A003-1; CAT assay kit; cat no. A007-1; total protein assay kit; cat no. A045-3; Nanjing Jiancheng Bio Co., Ltd.). The levels of MDA, GSH-Px, SOD and CAT were normalized to the content of total protein.
8
Hippocampal MDA and Catalase Assay
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The contents of malondialdehyde (MDA) and catalase (CAT) activity of the hippocampus were measured by lipid peroxidation MDA Assay Kit and CAT Assay Kit (Nanjing Jiancheng Bioengineering Institute, China), respectively, following the manufacturer’s instructions.
9
Zebrafish Liver CAT and MDA Assays
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CAT activity and MDA level of zebrafish liver were measured by using the CAT assay kit and MDA assay kit (Nanjing Jiancheng Bioengineering Institute, Nanjing, China) according to the instruction manual, respectively.
10
Antioxidant Activity Assay of EGCG in UVB-Irradiated HLE B-3 Cells
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HLE B-3 cells were seeded into 6-well plates (5.0×105 cells/well) overnight and were then exposed to UVB irradiation (30 mJ/cm2) in the presence or absence of EGCG pretreatment (50 µM) for 2 h, followed by a 24 h culture. At the specified time point, the cells were harvested, the cell pellets were collected by centrifugation at 400 × g for 5 min at 4°C, followed by suspension in PBS and ultrasonication on ice at moderate energy for 5 sec/time for a total of 10 min. Furthermore, the supernatants were collected after centrifugation at 5,000 × g for 10 min at 4°C. According to the manufacturer's instructions, the activities of catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), as well as the levels of GSH, H2O2 and hydroxyl free radicals were detected using the corresponding kits. The following kits were used (all purchased from Nanjing Jiancheng Bioengineering Institute): CAT assay kit (cat. no. A007-1-1), SOD assay kit (cat. no. A001-3-2), GSH-Px assay kit (cat. no. A005-1-2), reduced GSH assay kit (cat. no. A006-2-1), hydrogen peroxide assay kit (cat. no. A064-1-1), hydroxyl free radical assay kit (cat. no. A018-1-1). Finally, the absorbance was measured at a suitable wavelength (Multimode Plate Reader; PerkinElmer, Inc.).
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