Serum oxidized LDL was analyzed using an oxLDL ELISA kit according to the manufacturer's recommendations. Absorbances were read using a BioTeK Synergy H1 Hybrid Reader (BioTek Instruments Inc., Winooski, VT, USA) at the recommended wavelength (450 nm), and results were analyzed on https://www.myassays.com using linear regression (R2 = 0.9989, y = 0.1258x − 0.0041).
Biotek synergy h1 hybrid reader
Manufactured by Agilent Technologies
Sourced in United States
The BioTeK Synergy H1 Hybrid Reader is a multi-mode microplate reader that can perform absorbance, fluorescence, and luminescence detection. It is designed for a variety of applications, including cell-based assays, enzyme-linked immunosorbent assays (ELISAs), and nucleic acid quantification.
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Lab products found in correlation
Facsaria 3 cell sorter, by BD (1 mentions)
Jem 2100, by JEOL (1 mentions)
Agilent 1260 infinity hplc system, by Agilent Technologies (1 mentions)
Fl 4600 fluorescence spectrometer, by Hitachi (1 mentions)
Fluoview fv3000 confocal microscope, by Olympus (1 mentions)
Glucometer, by Roche (1 mentions)
Live dead baclight bacterial viability kit, by Thermo Fisher Scientific (1 mentions)
30 protocols using biotek synergy h1 hybrid reader
1
Quantification of Oxidized LDL
2
Serum Biomarker Measurement by ELISA
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Serum from blood collected in plain tubes was used for measurements of RBP4, adiponectin, and leptin using the respective ELISA kits according to the manufacturers’ instructions. Absorbances were read on BioTeK Synergy H1 Hybrid Reader (BioTek Instruments Inc., Winooski, VT, USA) at 450. The results were analyzed on www.myassays.com using four parametric test curve; adiponectin (R2 = 0.9954), RBP4 (R2 = 9969), leptin (R2 = 0.9916).
3
Platelet Adhesion to Extracellular Matrix
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Adhesion of platelets to laminin and collagen was determined according to the described procedures with minor modifications [38 (link), 39 ]. Platelets were pre-incubated with policosanol extract at various concentrations (10 min at 37 °C) and DMSO served as vehicle control. Using a 96-well plate, 50 μL of 40 μg/mL of collagen (0.05 % in CH3COOH) or 50 μL of laminin solution (1 mg/mL in phosphate buffer solution) was pre-incubated for 2 h. The wells were subsequently treated with 200 μL of PBS containing 1 % BSA for 1 h after aspiration and washing with 200 μL of PBS. AA (0.5 mM), ADP (10 μM), and collagen (5 μg/mL) were used as platelet activators, and incubated with 50 μL of platelet suspension per each coated well at 37 °C for 1 h. The plate was washed at least three times with 200 μL PBS to remove unattached platelets. Subsequently, 140 μL of the substrate solution containing 1 mg/ml p-nitrophenyl phosphate in citrate buffer (0.1 M sodium citrate, 0.1 M acetic acid and 0.1 % (w/v) Triton X-100, pH 5.4) was added to each well. The reaction was stopped after 1 h incubation at 25 °C and the color was developed by addition of 100 μL of NaOH (2 N). The absorbance of the reaction product, p-nitrophenol, was measured at 405 nm using BioTeK Synergy H1 Hybrid Reader (BioTek Instruments Inc., Winooski, VT, USA).
4
Measurement of Serum F2-Isoprostanes by ELISA
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Serum from blood collected in plain tubes was used for measurements of Serum F2-isoprostane using the respective ELISA kits according to the manufacturers' instructions. Absorbances were read on BioTeK Synergy H1 Hybrid Reader (BioTek Instruments Inc., Winooski, VT, USA) at the appropriate wavelengths (450 nm). The results were analyzed on http://www.myassays.com/ using four-parametric test curve; F2-isoprostane (R2 = 1).
5
Quantification of Serum Biomarkers
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Serum from blood collected in plain tubes was used for measurements of adiponectin, leptin, F2-isoprostane, and insulin using the respective ELISA kits according to the manufacturers' instructions. Absorbance was read on BioTeK Synergy H1 Hybrid Reader (BioTek Instruments Inc., Winooski, VT, USA) at the appropriate wavelengths (450 nm for insulin, leptin, and F2-isoproatane and 450 and 590 for adiponectin). The results were analyzed on http://www.myassays.com/ using four parametric test curve: adiponectin (R2 = 0.9914), insulin (R2 = 1), leptin (R2 = 0.9996), and F2-isoprostane (R2 = 1).
6
Liver Antioxidant and Insulin Assessment
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For these assays, 100 mg of liver samples was homogenized in 1 ml cold phosphate‐buffered saline (1× PBS, pH 7.4), centrifuged at 5323 g for 5 min, and the supernatant was collected. Serum insulin, liver F2‐isoprostane, and total antioxidant status were quantified using ELISA kits (Elabscience Biotechnology Co. Ltd, Wuhan, China) according to the manufacturer's instructions. The absorbance was read at 450 nm on a BioTeK Synergy H1 Hybrid Reader (BioTek Instruments Inc., Winooski, VT, USA).
7
Characterization of ZGO:Mn Nanomaterials
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The morphology of ZGO:Mn nano verifier was measured by a transmission electron microscope (TEM) (JEOL, JEM-2100, Japan) with a working voltage of 200 kV. Powder X-ray diffraction (XRD) patterns of ZGO:Mn nano verifier was measured on an X-ray diffractometer (Burker, D8 Advance, Germany) with Cu-Kα radiation (λ = 1.5406 Å) to determine the crystal structure. Absorbance was measured and recorded on a BioTeK Synergy H1 Hybrid Reader (BioTek, USA). Persistent luminescence performance of ZGO:Mn nano verifier was measured on a Hitachi FL4600 fluorescence spectrometer (Hitachi, Japan). The morphologies of S. putrefaciens and R. palustris were observed with a field emission scanning electron microscope (SEM) (Zeiss, SIGMA,Germany). Microbial consortia were imaged on the Olympus Fluoview FV3000 confocal microscope (Olympus, Japan). Current intensity of microbial cells was measured by a digital source meter (Keysight B1500A, Germany) connected to a probe station (PRCBE LAB, China). Fluorescence-activated cell sorting (FACS) was performed by BD FACSAria III Cell Sorter (BD Biosciences, USA). The Agilent 1260 Infinity HPLC system (Agilent, USA) was used to measure the concentration of lycopene.
8
Liver Tissue TBARS Quantification
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Liver tissue TBARS was measured as reported previously with some modifications [9 (link)]. Briefly, liver tissue (80 mg) was homogenized in 250 uL of 0.25Hcl and mixed with 250 uL of 0.375 % thiobarbituric acid and 250 uL of 15 % tricholoroacetic acid. Then, the mixture was incubated at 100 °C for 10 min and cooled before centrifuging at 3000 rpm for 15 min. Finally, absorbance of 100 uL of each sample and standard (TMP) were measured on BioTeK Synergy H1 Hybrid Reader (BioTek Instruments Inc., Winooski, VT, USA) at 532 nm, and the results expressed as uM MDA/mg tissue.
9
Platelet Aggregation Assay with ADP
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Platelet aggregation study was done using ADP as agonist according to Vaiyapuri and Gibbins [14 ] with minor modifications. Rat whole blood was collected into a vacutainer tube containing 3.2% sodium citrate (9 : 1, v/v) and mixed thoroughly by inverting the tubes for several times. The blood was centrifuged at 100 ×g for 20 minutes to obtain platelet rich plasma (PRP). 50 ng/mL of prostacyclin was added to prevent platelet activation. PRP was centrifuged again at 240 ×g for 10 minutes to sediment platelet pellet. The platelet pellet was resuspended in Tyrode-HEPES buffer and adjusted to a cell density of 108 cells/mL. BioTeK Synergy H1 Hybrid Reader (BioTek Instruments Inc., Winooski, VT, USA) was used as analytical tool. 100 μL of platelet cells was loaded into each well and 1 μL of ADP was added in. The optical densities at 405 nm were read for 20 minutes. Reading was taken at a fixed time interval of 1 minute. The plate was shaken in double orbital mode at a frequency of 282 cpm, 3 mm.
10
Cytokine Quantification by ELISA
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Serum CRP, IL-6 and TNF-α were measured using the respective ELISA kits according to the manufacturers’ instructions. Absorbance was read on BioTeK Synergy H1 Hybrid Reader (BioTek Instruments Inc., Winooski, VT, USA) at 450 nm. Results were computed into fold changes by dividing the results of the different groups with those of the normal group.
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