For the evaluation of renal oxidative stress, we measured MDA levels and the GSH content. The MDA level was assessed colorimetrically with the BioVision kit (Catalog# K739-100, Milpitas Boulevard, Milpitas, USA) that is based on the reaction of MDA in the sample with thiobarbituric acid (TBA) to generate the MDA-TBA adduct which can be easily quantified colorimetrically (OD 532 nm). The GSH content was determined colorimetrically using the BioVision kit (Catalog# K464-100, Milpitas Boulevard, Milpitas, USA) based on an enzymatic cycling method in the presence of GSH and a chromophore. The reduction of the chromophore produces a stable product, which can be followed kinetically at 450 nm. The procedures were conducted in accordance with the manufacturer's instructions to ensure accuracy and reliability.
Biovision kit
Manufactured by Abcam
Sourced in United States
The BioVision kit is a laboratory reagent used for conducting biochemical and cell-based assays. It provides the necessary components for performing various analyses, including colorimetric, fluorometric, and luminometric measurements. The kit's core function is to facilitate the quantification and detection of analytes, such as enzymes, metabolites, and other biomolecules, in a variety of sample types.
Automatically generated - may contain errors
26 protocols using biovision kit
1
Evaluation of Renal Oxidative Stress
2
Renal Function Assessment: BUN and Creatinine
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To assess the renal function, we measured serum BUN and creatinine levels using commercially available kits. BUN levels were determined colorimetrically with the BioVision kit (Catalog# K375-100, Milpitas Boulevard, Milpitas, USA). The principle of this is a kit comprised of the hydrolysis of urea by urease to produce ammonia and CO2. The ammonia that is produced then combines with hypochlorite and a phenolic chromogen to generate a complex that is green in color. The color intensity that develops is closely correlated with the urea content of the sample and vice versa. Besides, creatinine levels were measured colorimetrically using the BioVision kit (Catalog# E4370-100, Milpitas Boulevard, Milpitas, USA). The principle of this kit comprised the modified kinetic Jaffe methodology for the determination of creatinine involving a protein-free filtrate and a reaction with picric acid in an alkaline solution, a surfactant, and other ingredients to minimize protein and carbohydrate interferences [32 (link)]. The assays were performed following the manufacturer's instructions.
3
Oxidative Stress and Antioxidant Markers in Tissues
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Serum Malondialdehyde (MDA): was estimated using colorimetric kits supplied from (Randox Company – Germany) [21 (link)].
[Total antioxidant capacity (TAC)] level: It was estimated in both kidney and brain tissues according to manufacturer's instructions using Randox kits [22 (link)].
[Acetylcholinesterase]: It was estimated in brain tissue. Acetylcholinesterase activity is carried out based on Ellman's method using Biovision kits [USA, CA 95035].
[Total antioxidant capacity (TAC)] level: It was estimated in both kidney and brain tissues according to manufacturer's instructions using Randox kits [22 (link)].
[Acetylcholinesterase]: It was estimated in brain tissue. Acetylcholinesterase activity is carried out based on Ellman's method using Biovision kits [USA, CA 95035].
4
Enzymatic Assay of GLO1 and GLO2
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Enzymatic activity assays of GLO1 and GLO2 were determined using BioVision kits (#k591 for GLO1 and #k490 for GLO2). After drug treatment, cell lysates were prepared using ice-cold assay buffer containing protease inhibitors and kept on ice for 10 min. Samples were centrifuged at 13,200 g at 4 °C for 10 min. Supernatants were collected and kept on ice for the enzymatic assay as following the manufacturers’ instructions.
5
Serum LDH Activity Determination
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The blood samples collected from all animals were centrifuged, serum separated and used to determine LDH activity using BioVision kit (Milpitas, CA, USA).
6
Assessing Honey Bee Selenate Stress
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This test was conducted to assess the honey bee physiological stress induced by exposure to Se. The level of H2O2 was quantified using the biological liquid of whole bee samples exposed to four selenate and selenite concentrations (0, 6, 60, 600) μg/mL. Bees were fed these concentrations in vitro through 1 M sugar syrup for 2 days while control bees were only administrated 1 M sugar syrup. Bees were individually crushed in 1.5 mL tubes with 300 μL ultra-sterilized water and centrifuged at 11,000g for 3 min. In order to eliminate the proteins, the supernatant containing the biological liquid was filtrated through a 10 kDa filter and the assay was conducted using BioVision Kit (CA, USA) as per the manufacturer’s instructions.
7
Enzymatic Creatinine Quantification
8
Serum and Liver FFA Determination
9
Gill Lactate Concentration Assay
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Gills were homogenized (TissueLyser, Qiagen, Germany) in lactate assay buffer for 1 min and centrifuged at 10,000 × g for 10 min at 4°C. The supernatant was collected for testing. The gill lactate concentration was determined with a BioVision kit (Lactate colorimetric assay kit II, BioVision, Milpitas, CA, United States).
10
Measuring Cellular Reactive Oxygen Species
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Reactive oxygen species (ROS) were detected using the Biovision kit (K936-250) (Milpitas, CA, USA) according to manufacturer’s protocol. Briefly, 1 × 104 cells per well were seeded in a 96-well plate. The next day, media was removed, and adherent cells were washed in 100 μL of ROS Assay Buffer. They were then incubated in 100 μL of 1× ROS Label diluted in ROS Assay Buffer for 40 min at 37 °C in the dark followed by treatment with 100 μL medium containing 0, 10, or 100 nM YM155 for 1 h. Fluorescence intensity was measured at Ex/Em = 495/529 nm, and data were reported as the change in fluorescence after background subtraction.
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