G6PD activity was assayed spectrophotometrically by monitoring the reduction of NADP+ at 340 nm by using a UV-2700 UV–VIS spectrophotometer (Shimadzu) as previously described [25] (link). The standard reaction mixture contained 20 mM Tris-HCl (pH 8.0), 0.01 M MgCl2, 200 μM NADP+, and 500 μM G6P. The reaction was initiated with the addition of the enzyme. Steady-state kinetic parameters were obtained by varying the concentration of one substrate (1–500 μM for NADP+ and 5–500 μM for G6P) and fixing the concentration of the second substrate at saturating concentration (200 μM for NADP+ and 500 μM for G6P). The experiment was performed in triplicate. The initial velocity obtained from the spectrophotometer was used to calculate the rate of product formation and was expressed as micromole of NADPH produced per minute per milligram of protein (μmole/min/mg), as calculated using the extinction coefficient of NADPH at 340 nm (6220 M−1 cm−1). Steady-state kinetic parameters, KM, kcat, and Vmax, were determined by fitting the data to the Michaelis–Menten equation by using GraphPad Prism (GraphPad Software).
Uv 2700 uv vis spectrophotometer
Manufactured by Shimadzu
Sourced in Japan
The UV-2700 UV-VIS spectrophotometer is a laboratory instrument manufactured by Shimadzu. The core function of the UV-2700 is to measure the absorbance or transmittance of a sample across the ultraviolet and visible light spectrum.
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Lab products found in correlation
F 4600 fluorescence spectrophotometer, by Hitachi (3 mentions)
F 7000 fluorescence spectrophotometer, by Hitachi (2 mentions)
Quartz cuvette, by Hellma (2 mentions)
Jem 1230, by JEOL (2 mentions)
Vector 22 ft ir spectrophotometer, by Bruker (2 mentions)
Mpeg sh5k, by Creative PEGWorks (1 mentions)
Zetasizer nano zs90, by Malvern Panalytical (1 mentions)
Hydrofluoric acid, by Sinopharm (1 mentions)
D8 advance instrument, by Bruker (1 mentions)
Analytical grade ethanol, by Sinopharm (1 mentions)
Vector 22 spectrometer, by Bruker (1 mentions)
Jem 2100 transmission electron microscope, by JEOL (1 mentions)
Cm300, by Philips (1 mentions)
Iraffinity 1s spectrophotometer, by Shimadzu (1 mentions)
Scanning electron microscope, by JEOL (1 mentions)
F 7000 spectrometer, by Hitachi (1 mentions)
Av 600 mhz, by Bruker (1 mentions)
Microtof q 2 mass spectrometer, by Bruker (1 mentions)
J 1500, by Jasco (1 mentions)
D8 advance, by Bruker (1 mentions)
34 protocols using uv 2700 uv vis spectrophotometer
1
Spectrophotometric Assay of G6PD Activity
2
Surface Modification of Gold Nanoparticles
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i-colloid™ Au 15 nm (optical density 1 [OD1]) and 20 nm (OD1) from IMRA America, Inc., were used to take advantage of its novel surface properties allowing sequential surface modification.26 –28 (link) Methoxy-PEG-thiol, MW 5k (methoxy-PEG-thiol, MW 5,345, substitution purity: >90%, abbreviated as “mPEG-SH5k”), was purchased from Creative PEGWorks (Chapel Hill, NC, USA). Fluorescein isothiocyanate-PEG-thiol, MW 5k (FITC-PEG-SH, substitution purity: >80%, “FITC-PEG-SH5k”), was purchased from Nanocs, Inc (New York, NY, USA). Cysteine-modified (RGD)4 peptide (MW: 1,845.98, purity: >95%, abbreviated as “RGD peptide”) with amino acid sequence RGDRGDRGDRGDPGC was purchased from RS synthesis, LLC (Louisville, KY, USA). Methoxy-PEG-SH5k, FITC-PEG-SH5k and RGD peptide were in powder form and dissolved in deionized (DI) water having an electric conductivity of <0.1 μS/cm without further purification. All solutions were freshly made as needed and used within 12 h. Dynamic light scattering (DLS) measurement with Zetasizer Nano ZS90 (Malvern Instruments, Malvern, Worcestershire, UK) was employed to measure an increase in hydrodynamic diameter of AuNPs with surface modification. UV-2700 UV-Vis Spectrophotometer (Shimadzu Corporation, Kyoto, Japan) was used for optical density (OD) or absorbance measurement of the colloidal suspension.
3
Characterization of N-Vinylcarbazole-Based Materials
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N-vinylcarbazole (98%), HSiCl3 (99%), and mesitylene (97%) were purchased from Aladdin Reagent Co., Ltd. (Shanghai, China). Analytical-grade ethanol (99.5%) and hydrofluoric acid (40% aqueous solution) were received from Sinopharm Chemical Reagent Co., Ltd. (SCRC; Shanghai, China). All reagents were used as purchased without further purification. The XRD spectrum was performed on a Bruker D8 Advance instrument (Bruker AXS GmbH, Karlsruhe, Germany) with Cu Kα radiation (λ = 1.5418 Å). TEM images were obtained on a JEM-2100 transmission electron microscope with an acceleration voltage of 200 kV (JEOL, Ltd., Akishima, Tokyo, Japan). The FTIR spectra were measured by a Bruker VECTOR 22 spectrometer (Bruker, Germany) with KBr pellets. The PL and excitation spectra were collected by a Hitachi F-4600 fluorescence spectrophotometer (Hitachi, Ltd., Chiyoda-ku, Japan). The UV-vis absorption spectra were measured by a Shimadzu UV-2700 UV-vis spectrophotometer (Shimadzu Corporation, Kyoto, Japan). The PL lifetime was obtained on a Zolix Omni-λ 300 fluorescence spectrophotometer (Zolix Instruments Co., Ltd., Beijing, China).
4
Characterization of Nanoparticle-Encapsulated E6
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The particle sizes and zeta potential of PION@E6 were measured using a Brookhaven-zeta plus Particle Analysis Device (Brookhaven Instruments Corporation, NY, USA) and a Zeta Potential Device (Nanjing Fuxin Analysis, China), respectively. Morphological characteristics of PION@ E6 were observed and photographed by Philips CM300 transmission electron microscopy (TEM) images. Analysis of ultraviolet-visible-near-infrared absorbance spectra and fluorescent spectra of free E6 and PION@E6 were carried out using a UV-2700 UV-VIS Spectrophotometer (Shimadzu, Kyoto, Japan) and an F-7000 Fluorescence Spectrophotometer (Hitachi Ltd., Tokyo, Japan), respectively. In addition, the stability of PION@E6 dissolved in deionized water was determined and further aggregation of PION@E6 was tested by colloid stability test after 5 weeks.
5
UV-Vis Analysis of DPP Compounds
6
Comprehensive Characterization of Nanomaterials
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UV-vis absorption spectra were measured via a Shimadzu UV-2700 UV-vis spectrophotometer. The transmission electron microscopy (TEM) images were obtained on a ThermoFisher Tecnai G2 Spirit TEM microscope. The Fourier transform infrared (FTIR) spectra were recorded on a Shimadzu IRAffinity-1S spectrophotometer. The dynamic light scattering (DLS) images were generated on a Malvern ZEN3700 DLS and Zeta Sizer. The Raman spectra were measured by a Renishaw's inVia Raman spectrometer.
7
Chlorophyll Extraction and Quantification
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Chlorophyll was extracted from the leaves of the NtSPS1 OE and WT tobacco plants at five stages (0, 3, 6, 9, and 12 DAS) according to the procedure described by Yan et al. [29 (link)]. The absorbance of the filtered extracts at 645 nm (D645) and 663 nm (D663) was determined using a UV-2700 UV-VIS spectrophotometer (Shimadzu, Tokyo, Japan). Chlorophyll a (CA) and chlorophyll b (CB) content was then calculated using the following equations: CA = 0.125 × (13.7 × (D663) − 5.76 × (D645)) and CB = 0.125 × (25.8 × (D645) − 7.6 × (D663)).
8
Rifapentine-Loaded PLA Microsphere Characterization
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Ultraviolet spectrophotometry was used to measure different concentrations of rifapentine and the standard curve was drawn. Then, 5 mg of RPSMs were weighed, placed in 5 mL of dichloromethane, sonicated for 20 minutes, centrifuged at 800 × g, and the supernatant was collected. The absorbance was measured and converted into a standard curve equation to obtain the drug concentration. The encapsulation and drug loading efficiency of three batches of prepared microspheres were measured three times and the average encapsulation and drug loading efficiency of the microspheres were calculated. The drug loading efficiency of RPSMs was calculated as the percent ratio of the actual drug content to the total microsphere weight, while the encapsulation efficiency was calculated as the percent ratio of the actual drug loading to the theoretical drug loading. The morphology of the microspheres was observed under a scanning electron microscope (JEOL, Tokyo, Japan). The Ultraviolet-visible (UV-vis) spectra of rifapentine, RPSMs and blank PLA microspheres at a concentration of 1 mg/mL were measured on a UV-2700 UV–vis spectrophotometer (Shimadzu, Japan).
9
UV-Vis Analysis of DPP Compounds
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UV-Vis measurements were carried out on a Shimadzu UV-2700 UV-VIS spectrophotometer. Samples for absorption measurements were contained in 1 cm × 1 cm quartz cuvettes with 1 cm light path (Hellma analytics). HEPES (20 mM, pH = 7.4, 150 mM NaCl) containing 1% Triton X-100 was used to make dilute solution of DPP1, DPP2 and DPP3 of 1.0 - 0.1 absorbance at 310 nm. Extinction coefficients were calculated by linear least-squares fitting of plots of A vs. concentration. All fits gave R2 values of ≥ 0.999.
10
Characterization of PION@E6 Nanoparticles
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The hydrate particle size was measured using a Brookhaven ZetaPlus Particle Analysis Device (Brookhaven Instruments Corporation, Holtsville, NY, USA). The zeta potential was measured using Zeta Potential Device (Nanjing Fuxin Analysis, Nanjing, China). The morphology and structure of PION@E6 were characterized by Philips CM300 transmission electron microscopy (TEM). UV-Vis/NIR spectra analysis of free E6 and PION@E6 were carried out using a UV-2700 UV-VIS spectrophotometer (Shimadzu Corporation, Kyoto Prefecture, Japan). Fluorescent spectra of free E6 and PION@E6 was analyzed by F-7000 Fluorescence Spectrophotometer (Hitachi, Tokyo, Japan). Absorption and fluorescence measurements were performed in 1×1 cm quartz cell at room temperature, and deionized water was used as a solvent. For spectral measurement, the obtained solution of PION@ E6 was dissolved 100 times in deionized water. Meanwhile, stock solution of E6 at the concentration of 1 mg/mL was prepared in 20% ethanol and further diluted in deionized water 100 times, and the final concentration of E6 was 10 µg/mL.
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