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Cary 300bio spectrometer

Manufactured by Agilent Technologies
Sourced in United States

The Cary 300Bio spectrometer is a UV-Vis spectrophotometer designed for biological applications. It measures the absorption of light by samples across the ultraviolet and visible light spectrum, providing data on the concentration and identity of molecules within the sample.

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4 protocols using cary 300bio spectrometer

1

Cyanobacteria Absorption and Fluorescence

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Fluorescence excitation and emission spectra of the cultures was measured with a PTI Quantamaster spectrometer (Horiba, Kyoto, Japan), using glass cuvettes. Two micromolars of DCMU were added, where indicated. For emission spectroscopy, excitation was set at 497 nm, unless otherwise indicated. Slit widths was fixed to 4 nm throughout the experiments. In vivo absorption spectra of highly scattering cyanobacterial cultures were measured using an integrating sphere fitted into the measuring chamber of the Quantamaster spectrometer (Fig. 2). This set‐up was also used to measure total absorption in Fig. 6. In vitro pigment spectra were measured using a Cary 300Bio spectrometer (Agilent Technologies Inc., Santa Clara, CA, USA; Fig. 2). PSII Quantum Yield (Fv/Fm) and fluorescence rise kinetics were measured with a Joliot‐type spectrophotometer (JTS‐10; Bio‐Logic, Grenoble, France).
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2

Quantifying Quinoa Husk Saponins

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The extraction yield of total saponins was measured following a previously described method, with modifications [28 (link)]. Firstly, 0.2 mL of quinoa husk extracts was evaporated to dryness in a 70 °C water bath and cooled down. Amounts of 0.4 mL of 5% vanillin-acetic acid solution and 1.6 mL of 72% perchloric acid were added in sequence and incubated in a 70 °C water bath for 10 min, followed by cooling with ice-cold water for another 10 min. Then, the mixture was mixed with 10 mL of 17 M acetic acid and incubated at 25 °C for 15 min. The absorbance was measured at 560 nm using a Cary 300 Bio spectrometer (Agilent Corporation, Santa Clara, CA, USA), with the oleanolic acid solution as standard and the sample solution as a blank control.
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3

Preparative Purification and Characterization

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Example 2

Medium pressure liquid chromatography was performed using an Isolera One system equipped with a dual wave-length UV detector from Biotage and Biotage SNAP Ultra columns. For purification via preparative TLC, silica gel plates (EMD, 1 mm, 20×20 cm) were used, which contained a fluorescence indicator F254. Column chromatography was carried out using silica gel 60, EMD Millipore. All solvents were purchased from EMD Millipore Corporation and used as obtained. Deionized water was purified by a Millipore Milli-Q filtering system equipped with one carbon and two ion-exchange stages. All mass spectral measurements were performed by an Agilent LC-TOF high resolution TOF analyzer at the University of California-Riverside. All NMR spectra were recorded on a Bruker Avance 500 MHz instrument. Residual solvent signals were used as a reference. All UV measurements were performed on a Cary 300 Bio spectrometer from Varian.

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4

Glutathione S-Transferase Activity Assay

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Gst activity was analyzed using Glutathione S-transferase Assay kit (Sigma) per manufacturer’s instructions with minor modification. YAMC cells treated with 4-HNE or sham were harvested in PBS and sonicated using a Sonic Dismembrator (Fisher Scientific, Pittsburgh, PA, USA). Cell lysates were centrifuged at 10,000 × g for 5 min and supernatants used for analysis. The reaction solution consisted of 490 μl Dulbecco’s PBS, 5 μl of 200 mM L-glutathione reduced, 5 μl of 100 mM 1-chloro-2,4-dinitrobenzene as substrate, and 1 μl of cell lysate. Absorbance was measured at 340 nm at 37 °C for 5 min using a Cary 300 Bio spectrometer (Varian, Santa Clara, CA, USA). Gst activity was normalized to milligram total protein.
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