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The GPX, GR and GST assays were performed using a Power Wave × 340 spectrophotometer (Bio-Tek Instruments Inc., software KC4 v3.0).
Protein concentration was determined spectrophotometrically at 280 nm using a molar extinction coefficient of 2.66 and 1.06 cmL1 mg 1g–1 for lysozyme and insulin, respectively, with a Power Wave Biotek Instruments spectrophotometer (Highland Park, Box 998, Winooski, VT, serial no. 143379).
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Absorbance was measured at 405 nm using a Biotek power wave XS 2 reader (Winooski, USA).
The levels of formazan formed were quantified using a Microplate Power Wave X Scanning Spectrophotometer (Bio-TEK Instruments, Inc. Winooski, VT, USA).
The plates were rested for five minutes and the absorbance readings taken at 540 nm with a Biotek power wave XS2 reader.
Finally, for the ELISA readout, the absorbance of each well at 450 nm wavelength (A450) was measured using a microplate reader (Power Wave XS, Bio-Tek).
A total of 100 μL of reaction liquid was transferred to a 96-well plate to measure the absorbance at 450 nm using a microplate reader (Power Wave XS2, Gene Company, USA).
The total protein concentration in an adipose tissue sample was measured at 562 nm using a Bio-Tek Power Wave XS (Bio-Tek Instruments, USA) spectrophotometer and bicinchoninic acid (BCA) test Protein Assay Reagent (Pierce, Beijerland, Holland) according to the manufacturer's instructions.
Absorbance readings were taken in a microplate spectrophotometer (Biotek- Power Wave XS2), or an OceanOptics USB 4000 for the tests with crocin, with magnetic stirring and Peltier heating.
The designers assert that the floor mouths of the horns are additionally loaded acoustically by a vertical wooden horn expansion and the room's ceiling to create a 10 Hz "full power" wave at the listening position.
The absorbance of the reaction mixtures before and after substrate incubation was measured at 405 nm on a micro-plate reader (Power Wave Biotek Instrument Inc , USA.
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