Glucose oxidase reagent

Manufactured by Merck Group

Glucose oxidase reagent is a laboratory product used to measure glucose levels. It is a biochemical assay that employs the enzyme glucose oxidase to quantify the concentration of glucose in a sample.

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Lab products found in correlation

S8ap0 stereomicroscope, by Leica (1 mentions) Tissue triglyceride assay kit, by Applygen (1 mentions) T20 cell counter, by Bio-Rad (1 mentions) Trehalase, by Merck Group (1 mentions)

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Glucose oxidase (336 citations) Glucose reagent (9 citations) Glucose oxidase-peroxidase reagent (2 citations)

4 protocols using glucose oxidase reagent

Glucose Uptake Measurement under Stress

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The glucose uptake from the media was analyzed by using the glucose oxidase reagent (Sigma G3660). Briefly, cells were differentiated using the above media (4500 mg/L of glucose) and then treated with stress agent and ginsenosides. The media was collected and incubated with glucose oxidase at 1 μM for 1.5 hours at 37°C and then with dianoisidine reagent at 37°C for 30 minutes. The absorbance was taken spectrophotometrically at 540 nm and the amount of glucose left in the medium is calculated from the standard glucose, thus giving the glucose utilization under stress and ginsenoside treatment.[19 (link)]

Ponnuraj S.P., Siraj F., Kang S., Noh H.Y., Min J.W., Kim Y.J, & Yang D.C. (2014). Amelioration of insulin resistance by Rk1 + Rg5 complex under endoplasmic reticulum stress conditions. Pharmacognosy Research, 6(4), 292-296.

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Developmental and Reproductive Traits of Nilaparvata lugens

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Females were allowed to lay eggs for 2 h, and the hatched nymphs were then monitored every 12 h. Newly hatched 1^st-instar nymphs (n = 20, 0–12 hAE) were collected, and each individual was raised separately in a glass tube. The developmental times of 1^st-, 2^nd-, 3^rd-, 4^th- and 5^th-instar stages were monitored every 12 h. Glucose and triglyceride levels were measured in pooled 24h-adult females (n = 15) as reported previously [46 (link)]. Glucose levels were measured using glucose oxidase reagent (Sigma-Aldrich) according to the manufacturer’s instructions. Triglyceride contents were quantified by enzymatic hydrolysis using the GPO Trinder method with a tissue triglyceride assay kit (Applygen Technologies), according to the manufacturer’s instructions. The glucose and triglyceride contents were calculated based on three biological replicates. Adult longevity was determined by recording the mortality of newly emerged adult females (0–3 hAE, n = 43 for WT^SW and n = 42 for NlInR2^E4) and males (0–3 hAE, n = 52 for WT^SW and n = 42 for NlInR2^E4) every 12 h. To determine fecundity, newly emerged adult females (0–12 hAE) were collected for paired mating assays. Each female (n = 20) was allowed to match with two males in a glass tube. The insects were removed 10 days later and the laid eggs were counted under a Leica S8AP0 stereomicroscope.

Xue W.H., Xu N., Chen S.J., Liu X.Y., Zhang J.L, & Xu H.J. (2021). Neofunctionalization of a second insulin receptor gene in the wing-dimorphic planthopper, Nilaparvata lugens. PLoS Genetics, 17(6), e1009653.

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In Vitro Glucose Uptake Assay in C2C12 Cells

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The in vitro glucose uptake assay was carried out by using a modified methods described by Van de Venter et al. (2008).^{27 (link)} The C2C12 cells were grown to 80-90% confluency and then dislodged by a brief exposure to 0.25% Trypsin in PBS, counted (25,000 cells/ml) using a T20 cell counter (Bio-Rad) and suspended in complete growth medium. The cells were then seeded in a 96 well plate in a total volume of 200 μl/well and incubated at 37°C until they were 80% confluent. The cells were then treated with the crude plant extracts to reach a final concentration of 500 μg/ml and 1 mg/ml. The cells were incubated for 1, 3 and 6 h at 37 ^oC and at each time point, 50 µl of the supernatant was transferred from each well to a new 96 well plate, where 100 µl of glucose oxidase reagent (Sigma) was added. The mixture was then incubated at 37°C for 15 min and the absorbance measured at 540 nm using a microplate spectrophotometer. Cells treated with insulin at concentrations of 100 nM were used as positive control for the measurement of glucose uptake, while the untreated cells were used as negative controls. Glucose uptake was calculated as in the formula below, and expressed as increase in glucose uptake compared to untreated controls %.

Kamga-Simo FD I.I.I., Kamatou G.P., Ssemakalu C, & Shai L.J. (2021). Cassia Abbreviata Enhances Glucose Uptake and Glucose Transporter 4 Translocation in C2C12 Mouse Skeletal Muscle Cells. Journal of Evidence-based Integrative Medicine, 26, 2515690X211006333.

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Quantifying Glycogen and Trehalose in Cells

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Glycogen and trehalose were measured as described by Parrou and François (1997) (link). Ten OD_600nm of cells were incubated in 250μl of 0.25M Na₂CO₃ at 95°C for 4 hr. After this the solution was adjusted to pH 5.2 with 150μl 1M acetic acid and 600 μl of 0.2M sodium acetate. For glycogen, samples were incubated overnight with 2 U/ml of amyloglucosidase isolated from Aspergillus niger (Sigma Aldrich) at 57°C with constant agitation. For trehalose, samples were incubated overnight with 0.05 U/ml of trehalase (Sigma Aldrich) at 37°C. The glucose liberated from either glycogen or trehalose was determined by incubating 50μl of sample with 500μl of glucose oxidase reagent (Sigma) for 30 min at 37°C. The reaction was terminated with 500μl of 12M H₂SO₄ and the absorbance measured at 420 nm.

Huang J., Mousley C.J., Dacquay L., Maitra N., Drin G., He C., Ridgway N.D., Tripathi A., Kennedy M., Kennedy B.K., Liu W., Baetz K., Polymenis M, & Bankaitis V.A. (2018). A Lipid Transfer Protein Signaling Axis Exerts Dual Control of Cell-Cycle and Membrane Trafficking Systems. Developmental cell, 44(3), 378-391.e5.

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