The FRAP assay was carried out according to the method of Maizura et al. [28 ] with some modifications. FRAP reagent was prepared by dissolving 10 mM TPTZ solution in 40 mM HCL and 20 mM iron (III) chloride solution in a proportion of 10 : 1 : 1 (v/v) with acetate buffer. The reagent was prepared daily and stored at 370°C. At 593 nm in a UV visible spectrophotometer (UV-VIS 1200, Shimadzu Corporation, Japan), absorbance was measured. Ascorbic acid was used as the standard.
Uv vis 1200
Manufactured by Shimadzu
Sourced in Japan
The UV–vis 1200 is a spectrophotometer designed for the measurement of absorbance, transmittance, and reflectance of samples in the ultraviolet and visible light regions of the spectrum. It features a wavelength range of 190 to 1100 nanometers and can be used for a variety of applications, including quantitative analysis, kinetic studies, and qualitative identification of compounds.
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Lab products found in correlation
4 protocols using uv vis 1200
1
Ferric Reducing Antioxidant Power Assay
2
Biomass, Pigment, and Protein Quantification
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Cell growth was determined by measuring the dry weight of the biomass. Cells were filtered using filter paper (GC-50, ADVANTEC), oven dried at 105 °C for 2 h, and were placed in a desiccator for 1 h before measuring the weight. Biomass weight was calculated by subtracting the dry weight of the blank. Chlorophyll a and phycocyanin were repeatedly extracted using 80 % acetone and 0.01 M potassium phosphate buffer of pH 7.8, respectively. Pigment contents was calculated by measuring their absorbance at 750 nm using spectrophotometer (UV–vis 1200, Shimadzu). Protein was extracted by salting out, and its concentration was determined by Lowry et al. (1951 (link)).
3
UVA Photostability Evaluation of Fatty Acids
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The tested group (LW2-FA, CW2-FA, CEG-FA), BSA-FA, and peptide/protein-only groups were irradiated with UVA using a UVA lamp (365 nm, UVL-56 Handheld UV Lamp, 6 W; UVP, USA) at the same time. The distance between the lamp and the samples was fixed at 20 cm. All the samples were measured at 319 nm (UV-vis 1200, Shimadzu, Japan) using 10 mm cuvettes with MOPS buffer as a blank. The contents of FA samples were checked at 0, 2, 4, 8, and 12 h. The reservation ratio (RR) of FA was determined according to the following formula:
where Abi is sample absorbance including FA at 319 nm as time, Abj is sample absorbance without FA at 319 nm as time, and Abo is the absorbance of the initial sample at the same wavelength. The UV-visible absorption spectra were scanned at the same time, in the range from 250 to 400 nm. All experiments were carried out in triplicate.
where Abi is sample absorbance including FA at 319 nm as time, Abj is sample absorbance without FA at 319 nm as time, and Abo is the absorbance of the initial sample at the same wavelength. The UV-visible absorption spectra were scanned at the same time, in the range from 250 to 400 nm. All experiments were carried out in triplicate.
4
Quantification of Photosynthetic Pigments and Paramylon
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Chlorophyll a, chlorophyll b and carotenoid content was estimated according to Lichtenthaler’s procedures (Lichtenthaler and Wellburn 1983 (link)). Pigments were extracted with 80% (v/v) acetone and the optical density was measured by a spectrophotometer (UV–vis 1200, Shimadzu, Japan) at wavelengths of 470, 645, and 663 nm.
Paramylon was extracted and purified according to Barsanti’s method (Barsanti et al. 2001 (link)). For sample preparation, 50 mL aliquots of cultures were centrifuged at 12,000 rpm for 30 min and harvested cells were washed three times with distilled water. Cell pellets were frozen at − 20 °C for 12 h to break the cells, and were then resuspended in a 1% (w/v) sodium dodecyl sulfate and 5% (w/v) Na2EDTA solution. The suspension was shaken for a while and incubated at 37 °C for 1 h. Paramylon granules were retrieved by centrifugation for 30 min at 12,000 rpm. The above extraction treatment was repeated until the supernatant was translucent. The obtained paramylon granules were then rinsed three times with 70 °C glass-distilled water and immediately collected on glass fiber filter paper (APFC type, Millipore).
Paramylon was extracted and purified according to Barsanti’s method (Barsanti et al. 2001 (link)). For sample preparation, 50 mL aliquots of cultures were centrifuged at 12,000 rpm for 30 min and harvested cells were washed three times with distilled water. Cell pellets were frozen at − 20 °C for 12 h to break the cells, and were then resuspended in a 1% (w/v) sodium dodecyl sulfate and 5% (w/v) Na2EDTA solution. The suspension was shaken for a while and incubated at 37 °C for 1 h. Paramylon granules were retrieved by centrifugation for 30 min at 12,000 rpm. The above extraction treatment was repeated until the supernatant was translucent. The obtained paramylon granules were then rinsed three times with 70 °C glass-distilled water and immediately collected on glass fiber filter paper (APFC type, Millipore).
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