Liver homogenate was prepared by grinding 0.5 g liver tissue in 4.5 g physiological saline. The levels of malondialdehyde (MDA) and superoxide dismutase (SOD) in the serum and liver tissue were investigated using a PowerWave 340 microplate reader (Bio-Tek Instruments, Inc., Winooski, VT, USA), according to the manufacturer instructions of the MDA (cat. no. 20130819) and SOD (cat. no. 20130517) kits (Nanjing Jiancheng Bioengineering Institute, Nanjing, China). Normal serum reference values: MDA, 3.46–3.79 nmol/ml; SOD, 136.4–149.1 U/ml.
Powerwave 340 microplate reader
Manufactured by Agilent Technologies
Sourced in United States
The PowerWave 340 is a microplate reader designed for absorbance-based assays. It can measure absorbance at multiple wavelengths within the range of 200 to 999 nm. The instrument is capable of reading 6- to 384-well microplates.
Automatically generated - may contain errors
6 protocols using powerwave 340 microplate reader
1
Liver Tissue MDA and SOD Analysis
2
Quantitative ELISA for IgG Antibodies
3
Rat Serum and Liver Lipid Analysis
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Following sacrifice, blood was collected from rat abdominal aortas and centrifuged at 206 × g for 15 min at 4°C to separate the serum. Serum TC, TG, HDL-C, LDL-C, ALT and AST levels were measured using a fully automatic blood biochemistry analyzer (TBA-40FR; Toshiba Medical Systems Corporation, Otawara, Japan) with commercial biochemical kits according to the manufacturer's protocol. Liver homogenate was prepared by grinding 0.5 g liver tissue in 4.5 ml absolute ethyl alcohol followed by centrifugation at 825 × g for 10 min and the supernatant was separated. Levels of TC, TG and TBA in the liver were measured using ELISA kits and a PowerWave 340 microplate reader (BioTek Instruments, Inc., Winooski, VT, USA), according to the manufacturer's protocol.
4
Serum and Liver Biochemical Analysis
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Blood samples were collected and left to stand for 30 min at room temperature. Then centrifugation was operated at 1,200 × g for 15 min at 4°C to separate the serum. Serum TC, TG, HDL-C, LDL-C, ALT, and AST levels were measured using a fully automatic blood biochemistry analyzer (TBA-40FR; Toshiba Medical Systems Corporation, Otawara, Japan). The liver homogenate was prepared by grinding 0.5 g of liver tissue and then centrifuged at 1,500 ×g for 10 min to separate the supernatant. Levels of TC, TG, and TBA in the liver were measured using ELISA kits and a PowerWave 340 microplate reader (BioTek Instruments, Inc., Winooski, VT, United States).
5
Quantitative Cell Viability Assay Using MTT
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Cell viability was evaluated using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) assay as previously described (12 (link)). Briefly, 1×104 cells/well were seeded in 96-well plates and cultured for 18~24 h to reach 90% confluency. Following attachment, cells were washed twice with PBS, then serum-free medium added. Both serum deprived and control (10% serum) cells were harvested at 0, 12, 24, 36, 48 and 72 h. At each time point, the cell culture supernatants were discarded and 20 µl MTT solution was added to each well (0.5 mg/ml; Sigma Aldrich; Merck KGaA, Darmstadt, Germany), then the cells were cultured for a further 4 h. The supernatants were then removed and 200 µl DMSO was added to each well, with slight agitation for 15 min. The absorbance at a wavelength of 490 nm was then detected using a PowerWave 340 Microplate Reader (Bio-Tek Instruments, Inc., Winooski, VT, USA), with 4 replicates used for each well and a mean value calculated.
6
Quantitative HPLC Analysis of Resveratrol
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HPLC analysis was carried out using Perkin Elmer HPLC (Series 200) with C-18 reverse phase Discovery column (4.6 mm ID × 150 mm L; Sigma Aldrich) for the determination of resveratrol content. BIOTEK® Powerwave 340 microplate reader was used for different anti-oxidant assays. Resveratrol (RESV), 2,2′-azino-bis (3-ethyl benzothiazoline-6-sulphonic acid (ABTS), Trolox and FC reagent was purchased from TCI (Japan); 2,2-diphenyl-1-picrylhydrazyl (DPPH), Streptomycin, Sodium sulphate (Na 2 SO 4 ), Aluminium chloride (AlCl 3 ), Ferric chloride (FeCl 3 ) from Hi-Media Labs Pvt. Ltd. (India). Methanol (MeOH), Acetonitrile (ACN) and Conc. Sulphuric acid (H 2 SO 4 ) from Merck, Millipore, (USA), a Nitrocellulose membrane (NCM) from GE Healthcare and Life Sciences (USA), Orthophosphoric acid (H 3 PO 4 ) from Hi-Media (India).
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