Mitochondrial ATP activity was assayed using an ATP bioluminescence assay kit (Beyotime Institute of Biotechnology, China), according to the manufacturer's instructions. Mitochondrial transmembrane potential (△Ψm) in HPMECs was measured as the manufacturer's direction. Briefly, 25 nmol/L TMRM dye (Molecular Probes, Invitrogen, USA) was added to ECM medium for 30 min and assessed via spectrophotometer (SpectraMax 5; Molecular Devices, Sunnyvale, CA, USA). HPMECs were treated with salmon sperm DNA (10 μg/mL) for 2 h and then mitochondrial ROS was detected using MitoSOX (Invitrogen, USA) according to the manufacturer's instructions and measured via spectrophotometer. MPO, MDA, SOD and total antioxidant capacity were measured by MPO assay kits (Nanjing Jiancheng Bioengineering Institute, Nanjing, China), MDA assay kits (Beyotime Institute of Biotechnology, China), SOD assay kits (Nanjing Jiancheng Bioengineering Institute, Nanjing, China) and a rapid ABTS method (Beyotime Institute of Biotechnology, China), respectively.
Mpo assay kit
Manufactured by Nanjing Jiancheng
Sourced in China
The MPO assay kit is a laboratory instrument designed to measure the activity of the enzyme myeloperoxidase (MPO) in biological samples. MPO is an important enzyme involved in the immune response and its levels can provide insights into various physiological and pathological conditions. The kit utilizes a colorimetric or fluorometric detection method to quantify MPO activity, allowing researchers to assess its role in different biological systems.
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Lab products found in correlation
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Bca protein assay kit, by Solarbio (2 mentions)
Bicinchoninic acid bca protein assay kit, by Beyotime (2 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Spectramax5, by Molecular Devices (1 mentions)
Sod assay kit, by Nanjing Jiancheng (1 mentions)
Rapid abts method, by Beyotime (1 mentions)
Mitosox, by Thermo Fisher Scientific (1 mentions)
Atp bioluminescence assay kit, by Beyotime (1 mentions)
Hmgb1 elisa kit, by R&D Systems (1 mentions)
Interleukin 6 (il 6), by Nanjing Jiancheng (1 mentions)
Il 1β, by Nanjing Jiancheng (1 mentions)
Il 1α, by Nanjing Jiancheng (1 mentions)
Lactulose, by Biosynth (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Bca protein assay kit, by CWBIO (1 mentions)
Elisas, by R&D Systems (1 mentions)
Alt and ast assay kit, by Nanjing Jiancheng (1 mentions)
Tnf α elisa kit, by ExCell Bio (1 mentions)
70 protocols using mpo assay kit
1
Mitochondrial Bioenergetics and Oxidative Stress Assay
2
Quantifying Lung Tissue MPO Activity
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Lung tissues were weighted and homogenized in cold PBS. The supernatants were collected and the MPO activity in lung tissues was detected by the MPO assay kits (Jiancheng Bioengineering Institute, Nanjing, China) according to the manufacturer's instructions.
3
Measurement of Inflammatory Markers
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TNF-a, IL-6, IL-1β, IL-1α, IL-10 ELISA kit and MPO assay kits were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). The HMGB1 ELISA kit was obtained from R&D Systems. The BALF cytokine level and lung tissue MPO activity were measured according to the manufacturer’s instruction.
4
Lactulose Derivatives for Immunomodulation
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Lactulose was purchased from Carbosynth (Carbosynth Ltd.). Neu5Acα2,3Lactulose, Neu5Acα2,6Lactulose, Kdnα2,3Lactulose, and Kdnα2,6Lactulose were synthesized as described previously by our group (Zeng et al., 2018 (link)(Zeng et al., , 2019)) . The ELISA kits for TNF-α, IL-1β, serum IL-6, serum IL-10, and IL-12 were purchased from Wuhan Youersheng Trading Co. Ltd. The total SOD, POD, AKP, and MPO assay kits were purchased from Nanjing Jiancheng Bioengineering Institute. Analytical-grade NaCl, HCl, Na 2 HPO 4 , KCl, and KH 2 PO 4 were obtained from Guangfu Science and Technology Development Co. Ltd.
A mouse peripheral blood lymphocyte isolation kit (TBD) was purchased from Beijing Ya Anda Biotechnology Co. Ltd. Anti-mouse CD3 fluorescein isothiocyanate (FITC) was purchased from Ebioscience Company. Alsever's Solution was purchased from Beijing Ji Mei Biotechnology Co. Ltd. Chickens and guinea pigs were provided by China Three Gorges University (Yichang, China). We purchased RPMI 1640 from Gibco. Bovine serum albumin (BSA) was purchased from Sigma-Aldrich Corp. Microfiltration membranes (75 μm) and fluorescent microspheres (505/515) were purchased from Invitrogen. Six-well culture plates and cell scrapers were purchased from Corning Inc.
A mouse peripheral blood lymphocyte isolation kit (TBD) was purchased from Beijing Ya Anda Biotechnology Co. Ltd. Anti-mouse CD3 fluorescein isothiocyanate (FITC) was purchased from Ebioscience Company. Alsever's Solution was purchased from Beijing Ji Mei Biotechnology Co. Ltd. Chickens and guinea pigs were provided by China Three Gorges University (Yichang, China). We purchased RPMI 1640 from Gibco. Bovine serum albumin (BSA) was purchased from Sigma-Aldrich Corp. Microfiltration membranes (75 μm) and fluorescent microspheres (505/515) were purchased from Invitrogen. Six-well culture plates and cell scrapers were purchased from Corning Inc.
5
Quantifying Colonic Inflammation via MPO
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Inflammatory cell (polymorphonuclear neutrophil) infiltration into colonic tissue was quantified by measuring MPO activity with an MPO assay kit (Nanjing Jiancheng Bio-engineering Institute, China), following the manufacturer's instructions. MPO activity was expressed as units per gram of total protein (U/g).
6
Quantifying Neutrophil Infiltration via MPO
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Myeloperoxidase (MPO) acts as a marker of neutrophil infiltration, and its activity was determined as described in published literature [41 (link)]. Colon tissue was homogenized with ice-cold 0.9% saline containing 1% protease inhibitor cocktail to obtain 10% tissue homogenates, and that was centrifuged at 12,000 g for 20 min at 4 °C for the measurement of MPO activity with the MPO assay kit (Nanjing Jiancheng, Nanjing, China).
7
Quantitative Neutrophil Infiltration Assay
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MPO activity can be measured spectrophotometrically with the MPO assay kit (Nanjing Jiancheng Bioengineering Institute, Nanjing, China), to evaluate neutrophil infiltration quantitatively. MPO activity was expressed as units per gram of total protein (U/g).19 (link)
8
Quantifying Lung Myeloperoxidase Activity
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The lung tissue was homogenized on ice in five volumes of PBS and centrifuged at 15,000× g at 4°C for 10 minutes. Myeloperoxidase (MPO) activity in the lung tissues was measured using a MPO Assay Kit (Nanjing Jiancheng) following the manufacturer’s instructions. The MPO activity results are expressed as unit per gram of lung tissue weight (U/g).
9
Myeloperoxidase Activity Assay
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MPO activity was assayed according to the MPO assay kit manufacturer’s instructions (Jiancheng BioEngineering, Nanjing, China). Briefly, MPO is an enzyme found in granulocytes. We assessed MPO activity using a kinetic assay in which H2O2 is degraded by the MPO released from the samples of colon [25] (link). The change in absorbance at 460 nm was measured with a Life Science UV/Vis Spectrophotometer DU 530 (Beckman Coulter, USA). The data are presented as units per milligram of tissue, where 1 unit is equal to the amount of MPO required to degrade 1 µmol of H2O2 per minute at 25°C.
10
Murine Bronchoalveolar Lavage Fluid Analysis
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Mice were anesthetized with pentobarbitone (50 mg/kg I.P.) and tracheal intubations were performed. Then 1 mL preheated sterile saline was administered into the lung and extracted three times via a tracheal catheter. BALF was collected and kept on ice and centrifuged at 1200 × g for 10 min at 4°C to remove cell debris. The pellets were resuspended in 50 μL PBS and stained with Wright-Geimsa (KeyGen Biotech Co., Nanjing, China). Total cells and neutrophils were counted with hemocytometer in a double-blind manner. Protein levels in the BALF supernatants were determined using bicinchoninic acid protein assay (BCA) kit (KeyGen Biotech Co., Nanjing, China). An aliquot of BALF supernatant were used to assay the TNF-α, IL-6 levels using the respective ELISA kits (R&D, Minneapolis, MN, USA). MPO activity was measured with MPO assay kit (Nanjing Jiancheng Bioengineering Institute, Nanjing, China). All assays were conducted under the manufacturer’s instructions.
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