Affinity purified primary antibodies mouse anti-PLVAP mAb clone PAL-E were labeled with Alexa 647 fluorophores (Life Technologies, Carlsbad, CA), as per the manufacturer’s instructions. The antibody-fluorophore conjugate was quality controlled for conjugation efficiency by spectroscopy (Varian Cary 50 BIO; Agilent Technologies) and for aggregation by dynamic light scattering (Malvern Zetasizer Nano-ZS; Malvern Instruments, Malvern, United Kingdom).
Varian cary 50 bio
Manufactured by Agilent Technologies
Sourced in United States
The Varian Cary 50 Bio is a UV-Visible spectrophotometer designed for use in biological applications. It features a wavelength range of 190-1100 nm and can be used for various types of spectroscopic analysis.
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Lab products found in correlation
Cary 60, by Agilent Technologies (2 mentions)
Malvern zetasizer nano z, by Malvern Panalytical (1 mentions)
Hydrogen tetrachloroaurate 3 trihydrate, by Merck Group (1 mentions)
S 4700, by Hitachi (1 mentions)
H 7650, by Hitachi (1 mentions)
Sodium carbonate, by Scharlab (1 mentions)
Whatman 4 filter paper, by Cytiva (1 mentions)
Folin ciocalteu reagent, by Merck Group (1 mentions)
Methanol, by Carlo Erba (1 mentions)
Flame photometer 128, by Systronics (1 mentions)
Colorimetric assay, by Merck Group (1 mentions)
14 protocols using varian cary 50 bio
1
Fluorescent Labeling of Anti-PLVAP Antibody
2
Spectrophotometric Triglyceride Quantification
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Cells were scraped and lysed. Lipids were extracted using the chloroform/methanol (2:1) method (37 (link)). TG content was quantified spectrophotometrically by using “Triglycerides liquid” kit (Sentinel, Milan, Italy) in a Varian Cary-50Bio spectrophotometer (Agilent, Milan, Italy). Values were normalized to protein content. Data are expressed as percent TG content relative to controls.
3
Spectrophotometric Kinetics of Ferrous Iron Oxidation
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Iron solutions were prepared by dissolving ferrous iron sulfate in deionized water, adjusted to pH 2.0 with hydrochloric acid, and then filtered through 0.2 µm Whatman Puradisc polyethersulfone sterile syringe filters from Cytiva. Typically, 5 µL injections of a stock ferrous sulfate solution are added to a 1 mL protein solution prepared in 20 mM Mops, pH 7.4, sitting in a quartz cuvette with a screw-on cap stock to achieve a desired iron/protein ratio. The Fe(II) oxidation kinetics were followed under constant stirring on a Varian Cary 50 Bio or Cary 60 spectrophotometers from Agilent Technologies at 305 nm, where the Fe(III) oxo(hydroxo) species absorbs.
4
Synthesis and Characterization of Gold Nanoparticles
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Hydrogen tetrachloroaurate(iii ) trihydrate (HAuCl4·3H2O), silver nitrate (AgNO3), trisodium citrate (Na3C6H5O7), ascorbic acid (AA), hydrochloric acid (HCl), 5,5-dithio-bis-(2-nitrobenzoic acid) (DTNB), dopamine hydrochloride ((HO)2C6H3CH2CH2NH2·HCl), purchased from Sigma-Aldrich. All reagents were of analytical grade and used as received. Deionized water (DI) was used in all synthesis. All glassware was precleaned by aqua regia and then rinsed thoroughly with DI water. The morphology of nanoparticles was examined by Transmission Electron Microscope (TEM, Hitachi, H7650, Japan) at a voltage of 100 kV. In order to prevent aggregation of nanoparticles on the copper grid in the drying process, samples were prepared in 1 mg ml−1 BSA solution for TEM measurements.41 (link) The mean size of nanoparticles was determined using Image J software (open source) and TEM images. UV-visible spectroscopy (Varian Cary 50 Bio, Agilent Technologies, USA) was used to obtain absorbance spectrum of the prepared colloidal nanoparticles. Samples were numbered S1–S8. Scanning Electron Microscope (SEM, Hitachi S 4700) was used to determine of aggregated nanoparticles on the substrate. The surface charge of nanoparticles was determined with Zetasizer (Malvern, Nanoseries Nano-ZS). Schematic of the preparation process of the sensing platform is shown in Fig. 1 .
5
Total Phenolic Content Quantification
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The total phenolic content of the samples (cinnamon oleoresins and white chocolates) was estimated according to the method described by Udayaprakash et al. (2015) . The diluted sample (200 µl) was added to 1 mL distilled water and 200 µL Folin-Ciocalteu reagent. The solution was maintained at room temperature for 6 min before the addition of 2.5 mL 7% Na2CO3 solution and 2.1 mL distilled water. In order to obtain a stable coloured complex, the solution was incubated for 90 min at room temperature (∼20 °C). The absorbance was measured at 760nm using a UVvisible spectrophotometer (Varian Cary 50 Bio, Agilent Technology). The total phenolic content was expressed as milligrams of epicatechin equivalents per gram of the plant extract (mg ECE/g extract).
6
Kinetic Study of Iron Oxidation in Ferritin
7
Spectrophotometric Analysis of Metronidazole
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The amount of metronidazole which passed the membranes in the penetration studies was analyzed spectrophotometrically using Varian Cary 50 Bio UV- visible spectrophotometer (Agilent Technologies). Detection wavelength was 320 nm and calibration curves of standard solutions (0.3–20.0 μg/mL metronidazole) were used to calculate the concentration in each assembled receptor sample.
8
Measuring FixABCX Electron Bifurcation
9
Optimization of Methylene Blue Removal
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The effects of pH (2–9), temperature (293.16–323.16 K), shaking (0–150 rpm), initial dye concentration (25–2500 mg L−1), contact time (5–1440 min) and biosorbent dosage (1–10 g L−1) for the MB removal were investigated. The respective conditions have been mentioned in the relevant figures.
The samples were withdrawn from each flask at 5 minutes interval till an hour and every hour upto 5 hours and the final reading was taken at 24 hours. The samples were centrifuged and MB concentration in the supernatant was determined at 665 nm using UV-Visible Spectrophotometer (Varian Cary-50 Bio, Varian Inc., USA). Except column experiment, all the experiments were carried out in duplicate with a working volume of 200 mL in 500 mL Erlenmeyer flasks at room temperature (27±2°C) and under static conditions unless specified otherwise.
The samples were withdrawn from each flask at 5 minutes interval till an hour and every hour upto 5 hours and the final reading was taken at 24 hours. The samples were centrifuged and MB concentration in the supernatant was determined at 665 nm using UV-Visible Spectrophotometer (Varian Cary-50 Bio, Varian Inc., USA). Except column experiment, all the experiments were carried out in duplicate with a working volume of 200 mL in 500 mL Erlenmeyer flasks at room temperature (27±2°C) and under static conditions unless specified otherwise.
10
Ligand Selectivity Profiling of CYP168A1
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To determine ligand selectivity of CYP168A1, UV–visible difference spectra were acquired on a Varian Cary 50 Bio UV–visible scanning spectrophotometer (Agilent) for various ligands, including antifungal azole compounds and fatty acids. Both sample and reference chambers contained 1 ml of 1 μM CYP168A1 in 100 mM potassium phosphate, pH 7.4. Prior to initiating the titration, a baseline was recorded between 350 and 500 nm. Aliquots of ligand stock solutions prepared by serial dilution in dimethyl sulfoxide (DMSO) were added to the sample cuvette, whereas equal volume of vehicle solvent was added to the reference cuvette to determine the difference spectrum at varying concentrations. The absolute changes in absorbance deriving from a minimum of triplicate titrations were plotted as a function of ligand concentration and fitted to the one binding site model using the GraphPad Prism software (version 9.0.0, GraphPad software).
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