Berberine (B3251) (purity > 98%) and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Glucosamine (purity > 99%) was purchased from the Beyotime Biotechnology Company (Nanjing, China). The Roswell Park Memorial Institute 1640 medium (RPMI-1640) growth medium was purchased from (Hyclone Co., Logan, UT, USA). Fetal bovine serum (FBS), penicillin, and streptomycin were purchased from Lonza Walkerrsville (GIBCO CO., Allendale, NJ, USA). The concentrations of ROS and H2O2 in the liver were measured using a mouse ROS ELISA kit (RD-RX20367-48T) and H2O2 ELISA kit (RD-RX21392-48T) (Beijing Ruida Henghui Technology Development Co., Ltd., Beijing, China). HepG2 cells were purchased from the American Type Culture Collection (ATCC). Antibodies against Clock (ab93804) and Bmal1 (ab93806) were purchased from Abcam (Abcam, Cambridge, MA, USA). The cell counting kit-8 (CCK8) assay kit was purchased from DoJinDo (DoJinDo ChemTech Lim, Kumamoto, Tokyo, Japan). The Glucose Oxidase Assay Kit was obtained from APPLYGEN (APPLYGEN, Beijing, China). The cellular ROS and H2O2 assay kit was purchased from Beyotime (Biotechnology, Nanjing, China). Machine-filtered, pure water (Milli-Q) was used.
Glucose oxidase assay kit
Manufactured by Applygen
Sourced in China
The Glucose oxidase assay kit is a laboratory tool used to quantify the presence and concentration of glucose in a given sample. The kit contains the necessary reagents and protocols to perform a colorimetric analysis, allowing for the accurate measurement of glucose levels.
Automatically generated - may contain errors
Lab products found in correlation
Ab93806, by Abcam (1 mentions)
Glucosamine, by Beyotime (1 mentions)
Berberine b3251, by Merck Group (1 mentions)
Cell counting kit 8 cck 8 assay kit, by Dojindo Laboratories (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Hepg2 cell, by American Type Culture Collection (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Curcumin, by Merck Group (1 mentions)
Dexamethasone (dex), by Merck Group (1 mentions)
Incucyte s3 platform, by Sartorius (1 mentions)
Non essential amino acids (neaa), by Thermo Fisher Scientific (1 mentions)
7 protocols using glucose oxidase assay kit
1
Berberine and Oxidative Stress Regulation
2
Paeonol Improves Insulin Resistance in Adipocytes
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The glucose uptake study was performed according to method (Adam et al., 2016 (link); Li et al., 2017 (link)). Briefly, the 3T3-L1 adipocytes were cultured in DMEM supplemented with 10% FBS and1% antibiotic solution (10,000 U/ml penicillin G, 10mg/ml streptomycin) in 5% CO2 at 37 ℃, and changed every 48 h. Up to more than 80% of the cells differentiated and matured. For this study insulin resistance in 3T3-L1 was induced by incubation with TNF-α (10 ng/ml) for 96 h, After that, the medium was removed and the cells were treated with paeonol and its metabolites and pioglitazone (Pio, positive control) at 10 μM for 24 h. The supernatant was detected by a glucose oxidase assay kit (Applygen, China). The cells were washed once with PBS (0.01 mol/L, pH 7.4). Then, 100 μl of 5 mg/ml MTT was added to each well, incubated at 37°C for 2.5 h in a 5% CO2 incubator, dissolved in 150 μl of DMSO, and measured at 490 nm for detecting cell viability. Cell viability was calculated according to this formula:
3
Palmitic Acid-Induced Insulin Resistance Model
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We used palmitic acid (PA) to establish an insulin resistance model as described previously.5 (link) A PA (Sigma-Aldrich Co., St Louis, MO, USA) stock solution (0.1 mol/L) was prepared in absolute ethanol, and a 10% BSA stock solution was prepared with pure water (note: BSA was used to deliver PA to the cells). BSA and PA were formulated at a ratio of 9:1 by volume to prepare the working PA solution. HepG2 cells were treated with serum-free medium containing 0.25 mmol/L of PA. The glucose concentration in the medium was determined by a glucose oxidase assay kit (Applygen, Beijing, China) at 0, 12, and 24 hours of treatment to evaluate establishment of the insulin resistance model.
4
Curcumin-Induced Adipocyte Differentiation
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Curcumin (purity over 99%), dexamethasone, 3-isobutyl-1-methylxanthine (IBMX), insulin, and annexin V-fluorescein isothiocyanate/propidium iodide (V-FITC/PI) apoptosis detection kits were purchased from Sigma-Aldrich (St Louis, MO, USA). Dulbecco’s modified eagle medium: Nutrient Mixture F-12 (DMEM/F12), fetal bovine serum (FBS), Type I collagenase, penicillin and streptomycin were obtained from Gibco (Waltham, MA, USA). Triglyceride (TG) determination kit, glucose oxidase assay kit and glucose-6-phosphate dehydrogenase (GPDH) activity measurement kit were gained from Applygen Technologies (Beijing, China). Cell counting kit-8 (CCK-8), lactate dehydrogenase (LDH) release kit and oil red O (ORO) were provided by Nanjing Jiancheng Bioengineering Insititute (Nanjing, China).
5
Glucose Consumption in B16F10 Cells
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Collected the culture medium of B16F10 cells before and after GSK126 treatment, and utilized the glucose oxidase assay kit (Applygen, Cat# E1010) to detect the glucose concentration in the culture medium. The glucose consumption levels were calculated as the difference between the initial and the remaining glucose levels in the cell culture media of B16F10 cells subjected to GSK126 treatment. In the meanwhile, cell confluence was monitored in IncuCyte S3 platform (Sartorius, Göttingen, Germany). All glucose consumption levels were normalized to cell confluence.
6
Establishing Insulin Resistance in HepG2 Cells
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HepG2 human hepatoma cells were purchased from the National Infrastructure of Cell Line Resource of China and stored at the Clinical Research Center of Hebei General Hospital. The cells were cultured in MEM (Hyclone) containing 10% foetal bovine serum (Hyclone), 1% non-essential amino acids (Gibco) and 1% streptomycin (Hyclone) in the presence of 5% CO2 at 37°C. When the cells reached approximately 80% confluence, they were treated with serum-free medium (CON group) or serum-free medium containing 0.25 mmol/L palmitic acid (PA) to establish insulin resistance (PA group). At 0, 12, and 24 h, the glucose concentration in the medium was measured using a glucose oxidase assay kit (Applygen) to evaluate model establishment.
7
Glucose Quantification by Oxidase Assay
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The glucose content was measured by the Glucose Oxidase Assay Kit (APPLYGEN, Beijing, China). The level of glucose was calculated in μmol/mg protein.
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