Blood samples were collected by venipuncture into 3-mL serum-separating tubes on day 1 (n = 28) (0–24 h), day 2–5 (n = 24) (48–120 h), and on day 7–10 (168–240 h) (n = 24) after aSAH event. Patient follow-up was not possible in patients who died (n = 4). NC were sampled at only one-time point and the sampling and processing were identical for every group. These tubes were centrifuged at 3000 rpm during 5 min for serum collection. Serum was then aliquoted and stored at − 80 °C for further analysis. Specific enzyme-linked immunosorbent assay (ELISA) kits were used for the measurement of IL-18, GSDMD, and TF according to manufactures’ instructions as available in Table e2 , Epoch 2 Microplate Spectrophotometer reader was used (BioTek). Inflammatory cytokine IL-1β concentration was measured using BD Human Inflammatory Cytokine CBA kit (551,811, Becton–Dickinson Biosciences), acquired by BD FACS-Calibur flow cytometer (Becton–Dickinson Biosciences) and analyzed by FCAP Array software (Becton–Dickinson Biosciences).
Epoch 2 microplate reader spectrophotometer
Manufactured by Agilent Technologies
Sourced in United States
The EPOCH/2 is a microplate reader and spectrophotometer from Agilent Technologies. It is designed for absorbance measurements in a range of biological and chemical applications. The instrument provides accurate and reliable data with its high-performance optics and detection system.
Automatically generated - may contain errors
Lab products found in correlation
Facscalibur flow cytometer, by BD (1 mentions)
Fcap array software, by BD (1 mentions)
Lambda 35 uv vis spectrophotometer, by PerkinElmer (1 mentions)
Prism 5, by GraphPad (1 mentions)
Zetasizer nano z, by Malvern Panalytical (1 mentions)
Acetonitrile, by Merck Group (1 mentions)
Silver nitrate, by Merck Group (1 mentions)
Sodium borohydride, by Merck Group (1 mentions)
Malathion, by Merck Group (1 mentions)
Chlorpyrifos, by Merck Group (1 mentions)
Phorate, by Merck Group (1 mentions)
7 protocols using epoch 2 microplate reader spectrophotometer
1
Serum Biomarker Dynamics After Aneurysmal SAH
2
Bacterial Growth Kinetics in Microplates
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The bacterial strains were grown overnight in LB or MOPS EZ-rich medium containing 0.2% glucose. Cultures were diluted to 6 × 106 cells/mL in their respective medium and samples were prepared in triplicate by mixing 50 µL of cells and 50 µL of fresh medium to obtain 3 × 106 cells/mL. Assay was performed in Microtest plate, 96-well, flat base, polystyrene, sterile (Sarstedt) and growth was monitored using Epoch 2 Microplate Spectrophotometer reader (BioTek) with the following settings: OD = 600 nm, Temperature = 37°C, Reading = every 10 min for 22 hr, Continuous shaking.
3
Bacterial Growth Kinetics Assay
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The bacterial strains were grown overnight in LB or MOPS EZ-rich medium containing 0.2% glucose. Cultures were diluted to 6 x 10 6 cells/mL in their respective medium and samples were prepared in triplicate by mixing 50 µL of cells and 50 µL of fresh media to obtain 3 x 10 6 cells/mL. Assay were performed in Microtest plate, 96-well, flat base, polystyrene, sterile (Sarstedt) and growth was monitored using Epoch 2 Microplate Spectrophotometer reader (BioTek) with the following settings: OD= 600 nm, Temperature= 37 °C, Reading= every 10 min for 22 h, Continuous shaking.
4
Spectroscopic Analysis of BlsA Protein
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Pure protein samples in 20 mM Tris-HCl (pH 8.0) containing 500 mM NaCl were analyzed with an EPOCH/2 microplate reader/spectrophotometer (BioTek) or a Perkin Elmer Lambda 35 UV/Vis spectrophotometer. Protein samples were kept in the dark at 4°C for dark-adapted BlsA samples (dBlsA) before blue-light illumination, with light intensities ranging between 20 and 200 μmol/m2/s, for 3 min at 22°C and read using a spectral scan of 1 nm. The spectral absorbance properties of all derivatives were recorded at least twice from two different purified protein samples. For dark state recovery kinetics, purified samples were exposed to blue light for 3 min at 22°C and spectral scans were recorded at 505 nm for 20 min.
5
Enzymatic Assay for Feruloyl-CoA Synthetase
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The enzymatic assays were adapted from previously described methods [47 ,48 (link)], the schematic reaction was included in S1 Fig . The reaction mixture (200 μL) contained 2.5 mM of MgCl2, 0.5 mM of FA, 2.0 mM of ATP, 0.4 mM of coenzyme A, 40 ng of purified FCS1 and 100 mM of potassium phosphate buffer pH 7.8. The activity assay was initiated by the addition of the enzyme and incubated at 37°C for 10 min. Following incubation, 150 μL of the reaction were used to read the absorbance at 345 nm due to formation of feruloyl-CoA (ε345nm = 1.9 x 104 M-1cm-1), using an Epoch2 Microplate Reader spectrophotometer (BioTek, Winooski, VT, USA).
For determination of kinetic parameters of FCS1, the above reaction was assayed with substrate concentrations ranging from 0.05 mM to 0.50 mM. Mathematical adjustments were made using the software Graph Pad Prism 5.0 (GraphPad Software) to calculate the parameters. The assays were performed in triplicate and at least three independent experiments were carried out.
For determination of kinetic parameters of FCS1, the above reaction was assayed with substrate concentrations ranging from 0.05 mM to 0.50 mM. Mathematical adjustments were made using the software Graph Pad Prism 5.0 (GraphPad Software) to calculate the parameters. The assays were performed in triplicate and at least three independent experiments were carried out.
6
Colorimetric Pesticide Detection Using Silver Nanoparticles
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Silver nitrate (>99%) and sodium borohydride (>99%) were purchased from Sigma Aldrich Pvt. Ltd. Double distilled water was used for preparing the solutions. Chlorpyrifos, phorate, malathion, and acetonitrile were also procured from Sigma Aldrich Pvt. Ltd.
UV-visible spectra were recorded using Biotek Epoch 2 microplate reader spectrophotometer. Particle size and zeta potential analyses were done on Malvern Zetasizer Nano ZS. Raman spectra were acquired with a 532 nm laser.
Gold and silver nanoparticles show a color change on interaction with pesticides. Silver nanoparticles show higher surface plasmon resonance (SPR) than gold nanoparticles because of the large difference in the energy levels of conduction bands. Silver metal is comparatively harder and less reflective than gold. The SPR of silver nanoparticles appears at a smaller wavelength, allowing it to undergo wide variation upon interaction with analyte molecules. These properties make silver nanoparticles more suitable than gold nanoparticles to study the colorimetric response of given organophosphate molecules.27 (link)
UV-visible spectra were recorded using Biotek Epoch 2 microplate reader spectrophotometer. Particle size and zeta potential analyses were done on Malvern Zetasizer Nano ZS. Raman spectra were acquired with a 532 nm laser.
Gold and silver nanoparticles show a color change on interaction with pesticides. Silver nanoparticles show higher surface plasmon resonance (SPR) than gold nanoparticles because of the large difference in the energy levels of conduction bands. Silver metal is comparatively harder and less reflective than gold. The SPR of silver nanoparticles appears at a smaller wavelength, allowing it to undergo wide variation upon interaction with analyte molecules. These properties make silver nanoparticles more suitable than gold nanoparticles to study the colorimetric response of given organophosphate molecules.27 (link)
7
DPPH Radical Scavenging Assay for Essential Oils
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This assay was carried out according to the modified method of Braca et al. 19) . A stock solution of each essential oil (4.0 mg/mL) was prepared in dimethyl sulfoxide (DMSO) . After 180 μL of 0.05 mM DPPH methanol solution was mixed with 20 μL of the sample solution at different concentrations, decreases of the absorbance of the reaction mixtures were monitored at 517 nm using a UV-Vis spectrophotometer (EPOCH 2 Microplate Reader Spectrophotometer, BioTek Instruments, Winooski, VT, USA) after incubation at room temperature for 30 min in the dark. The DPPH inhibition (％) was calculated as follows, ％ Inhibi-tion＝ ［ (Ablank-Asample) /Ablank］ ×100, where Ablank is the absorbance of the control reaction (containing all reagents except the test compound) , and Asample is the absorbance of the test compound-containing reaction mixture. The sample concentration providing 50％ inhibition (IC 50 ) was calculated by plotting the inhibition percentages against the concentrations of the sample. All tests were carried out in triplicate and IC 50 values were reported as means. Ascorbic acid was used as the positive antioxidant agent.
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