The rat was anesthetized with pentobarbital sodium (50 mg/kg, i.p.) and placed in a stereotaxic apparatus. A craniotomy over the right amygdala was performed and a guide cannula (outer diameter: 0.64 mm; inner diameter: 0.45 mm) was implanted on the dorsal margin of the CeA, using the following coordinates (in mm): 2.5 caudal to bregma; 4.0 lateral to midline; depth, 7.5. The cannula was fixed to the skull with dental acrylic. The behavioral tests were carried out 5 days after the surgery. For drug application, a microinjection tubing probe was inserted through the guide cannula so that the probe protruded by 1 mm. The probe was connected to a PE-10 polyethylene tube filled with dissolved drug solution. The drug solution was pushed into the brain tissue by injecting 1μl air into the polyethylene tube with a microsyringe. Then the polyethylene tube was heat sealed and the probe was left in place for 10 min before the behavioral tests. The following drugs were used: Tempol, baicalein, apocynin (Tocris). Drugs were dissolved in DMSO (30% in deionized water) and diluted in ACSF (containing [in mM] 117 NaCl, 4.7 KCl, 1.2 NaH2PO4, 2.5 CaCl2, 1.2 MgCl2, 25 NaHCO3 and 11 glucose) at 1:100 to their final concentration for injection into the amygdala.
Apocynin
Manufactured by Bio-Techne
Sourced in United States, United Kingdom
Apocynin is a chemical compound that is used in laboratory research. It functions as a nicotinamide adenine dinucleotide phosphate (NADPH) oxidase inhibitor, which means it can block the activity of this enzyme complex. Apocynin is often used in experiments to study the role of NADPH oxidase in various biological processes.
Automatically generated - may contain errors
Lab products found in correlation
Losartan, by Bio-Techne (3 mentions)
Rapamycin, by Cell Signaling Technology (2 mentions)
Human ang 2, by Merck Group (2 mentions)
U73122, by Bio-Techne (2 mentions)
Pd123319, by Bio-Techne (2 mentions)
Baicalein, by Bio-Techne (1 mentions)
Tempol, by Bio-Techne (1 mentions)
Adenosine triphosphate (atp), by Merck Group (1 mentions)
N acetyl cysteine (nac), by Merck Group (1 mentions)
Apyrase, by Merck Group (1 mentions)
Cox 2, by Cell Signaling Technology (1 mentions)
Fk506, by Bio-Techne (1 mentions)
Adpβs, by Merck Group (1 mentions)
α tubulin, by Abcam (1 mentions)
Ar c 118925xx, by Bio-Techne (1 mentions)
Aacocf3, by Bio-Techne (1 mentions)
Nf157, by Bio-Techne (1 mentions)
Atpγs, by Bio-Techne (1 mentions)
Nf546, by Bio-Techne (1 mentions)
Tnp atp, by Bio-Techne (1 mentions)
10 protocols using apocynin
1
Amygdalar Modulation of Oxidative Stress and Behavior
2
Molecular Mechanisms of G Protein-Coupled Receptor Signaling Regulation
3
Neutrophil-Mediated Cell Killing Assay
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MC38, B16F10, KPC, and Pan02 cells expressing luciferase were used as target cells. For neutrophil-mediated cell killing assay, target cells were cocultured for 24–48 hours with neutrophils at various E:T ratios (e.g., 20:1) in presence or absence of indicated stimulant. In ROS inhibition experiments, neutrophils were pretreated with 300 μM Apocynin (Tocris Biosciences) or 5 μM diphenyleneiodonium chloride (Tocris Biosciences) and then used for target cell killing. For NK cell and neutrophil combination experiments, target cells were cultured at a E:T ratio of 10:1 NK cells and 2:1 neutrophils or together for 48 hours. In experiments, where specified, supernatants from either control- or IL-36–treated neutrophils were added instead of neutrophils. Luciferase was assessed using Steadyglo (Promega). and luminescence was read using a luminometer (Envision, PerkinElmer).
4
Ang II-Induced Vascular Remodeling
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GF 109203X was purchased from Calbiochem. Ang II, losartan, Go 6976, apocynin, and ML-171 were obtained from Tocris. Primary antibodies against CaV3.2, CaV3.1 and Nox1 were purchased from Novus Biologicals. Duolink PLA detection kit and all other chemicals were obtained from Sigma Aldrich. Data are expressed as means ± S.E., and n indicates the number of cells or arteries or animals. Paired t-test was performed to compare the effects of a given condition/treatment on whole cell current or arterial diameter. P values ≤ 0.05 were considered statistically significant.
5
Intrathecal Injection of Neuroprotectants
6
Freund's Adjuvant-Induced Inflammation
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Freund’s incomplete adjuvant was purchased from Sigma-Aldrich (Milan, Italy); desiccated mycobacteria, M. tuberculosis H37Ra and M. butyricum, were from Difco laboratories (Detroit, U.S.A.); minocycline hydrochloride and apocynin were from Tocris Bioscience (Bristol, U.K.); IL-1β (60 pg/ml) was from R&D Systems (Minneapolis, U.S.A.).
7
Immunological Assay Protocol for Cell Cultures
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Apocynin, SP600125, c‐Jun peptide, IT901, and pristmerin were purchased from Tocris Bioscience (Abingdon, UK). Ascorbic acid, collagenase IV, DMSO, pentobarbitone, PHA, and TNF‐α were bought from Sigma‐Aldrich (St. Louis, MO, USA ) and butorPHAnol from Cayman (Ann Arbor, MI, USA). Amoxicillin was supplied by RuiYang (Shandong, PRC). Recombinant human IFN‐γ and TGF‐β1 were bought from PeproTech (Rocky Hill, NJ, USA). Dynabead magnetic beads M450 were bought from Invitrogen. Anti‐rabbit (RRID:AB_2313567) and anti‐mouse (RRID:AB_10015289) secondary antibodies were purchased from Jackson ImmunoResearch Laboratories (West Grove, PA, USA). Anti‐rabbit IgG‐FITC (RRID:AB_631744) and anti‐mouse IgG‐FITC (RRID:AB_631735) antibodies were bought from Santa Cruz Biotechnology (Dallas, TX , USA). Anti‐rat MHC I antibody (Abcam ab15681, RRID:AB_302030) detects mouse MHC Class I antigens of d and b haplotype; anti‐rabbit MHC I antibody (Abcam ab110645, RRID:AB_10859600) detects human HLA B; anti‐rat MHC II antibody (Abcam ab23990, RRID:AB_447796) detects I‐A region of the mouse MHC; and anti‐mouse MHC II antibody (Abcam ab55152, RRID:AB_94419) detects human MHC II β chain HLA‐DPB1. The primary antibodies are listed in Table 1 .
8
Cardiomyocyte Culture and Pharmacology
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Mouse atrial cardiomyocyte HL-1 cells were a gift from Dr. William Claycomb at Louisiana State University Medical Center and were cultured as described previously (18 (link), 23 (link)). Nebivolol was a gift from Forest Laboratories Inc. (New York) and rapamycin was purchased from Cell Signaling Technology (Boston, MA). Human Ang II was purchased from Sigma-Aldrich. Losartan (AT1R inhibitor), PD123319 (AT2R inhibitor), apocynin (NADPH Oxidase inhibitor) and U73122 (Phospholipase C inhibitor) were purchased from TOCRIS Biosciences.
9
Cardiomyocyte Culture and Pharmacology
10
Apocynin Attenuates Diabetic-Induced Muscle Dysfunction
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The animals were randomly divided into the following groups: normoglycemic rats (control), non-treated diabetic rats (diabetes), and diabetic rats treated with apocynin (diabetes + apocynin); n = 6 in each group. Diabetes was induced by a single streptozotocin (STZ) injection (45 mg/kg body weight) (Sigma-Aldrich, St Louis, MO, USA) that was freshly dissolved in a citrate buffer (0.5 M, pH 4.5), and control rats (normoglycemic) received a citrate buffer injection, intraperitoneally, instead of STZ. Treatment for eight weeks with apocynin (Tocris Bioscience) started 1 week after STZ, by intraperitoneal injection of 3 mg/kg/day. Animals in the control groups were managed similarly to those in the apocynin-treated group. Variations in body weight and FBG were evaluated every week. All groups were evaluated during the same period. Once treatment was complete, the animals of all groups were fasted for 8 h and euthanized using cervical dislocation. Skeletal muscle samples from EDL and soleus were immediately dissected from both the left and right hind limbs in all animals. Isometric tension measurements were performed using fresh muscles, and the remaining muscles were stored at −80 °C until processed by the glutathione assay, to measure ROS levels and to evaluate mRNA expression levels by real-time RT-qPCR.
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