Whole blood samples were collected from the jugular vein of calves in 5 mL plain Vacutainer tubes. The samples were centrifuged, for 15 min at 1500 g speed, and sera were then harvested and preserved at −20°C until used. Serum levels of creatinine and blood urea nitrogen (BUN) were measured through commercial test kits (Spinreact, Girona, Spain), using ultraviolet (UV) spectrophotometer (Optizen 3220 UV, Mecasys Co. Ltd., Korea).
Optizen 3220uv
Manufactured by Mecasys
The Optizen 3220UV is a UV-Vis spectrophotometer designed for laboratory use. It provides accurate and reliable absorbance measurements across a wide wavelength range, serving as a versatile tool for various analytical applications.
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Lab products found in correlation
Pt3100, by Kinematica (2 mentions)
Unity 500 mhz spectrometer, by Agilent Technologies (1 mentions)
Dm2500 microscope, by Leica (1 mentions)
Jem 2100, by JEOL (1 mentions)
Invia raman microscope system, by Renishaw (1 mentions)
Tnbsa, by Thermo Fisher Scientific (1 mentions)
No 1 filter paper, by Cytiva (1 mentions)
Rm 1000, by Renishaw (1 mentions)
Inspect f50, by Thermo Fisher Scientific (1 mentions)
Spectrum one ftir, by PerkinElmer (1 mentions)
27 protocols using optizen 3220uv
1
Serum Creatinine and BUN Analysis
2
Optimization of PHA mcl Production
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Conditions for Optimum PHA mcl Production
The nitrogen concentration of the nutrient-limited media was evaluated using different amounts of NH 4 Cl. For this study, the cells were first grown in nutrient broth, then transferred to nutrient-limited media containing 0 g, 0.25 g, 0.50 g, and 1.0 g/l of NH 4 Cl with 200 mg/l of naphthalene. The samples were collected after five days of incubation at 22°C with 180 rpm and then prepared as described above for GC analysis. The monomer concentration was determined by comparison to the abundance peak of the standard and groups were compared by one-way ANOVA with Tukey's multiple test correction.
The optimum carbon concentration, temperature, and pH were determined for the PHA mcl producing bacteria. Naphthalene and anthracene were added to nutrient-limited media at different concentrations (200, 400, 600, 800, 1,400, and 1,600 mg/l) to evaluate the tolerance of the strains at 22°C for 5 d. The optical density (OD) of the strains was determined by UV spectrometry (Optizen 3220UV, Mecasys, Korea). The optimum temperature and pH of the strains were determined based on the OD observed after cultivating the organisms at seven different temperatures (5°C, 15°C, 20°C, 25°C, 30°C, 37°C, and 40°C) and five different pHs (5, 6, 7, 8, and 9) in nutrient broth media for 24 h.
The nitrogen concentration of the nutrient-limited media was evaluated using different amounts of NH 4 Cl. For this study, the cells were first grown in nutrient broth, then transferred to nutrient-limited media containing 0 g, 0.25 g, 0.50 g, and 1.0 g/l of NH 4 Cl with 200 mg/l of naphthalene. The samples were collected after five days of incubation at 22°C with 180 rpm and then prepared as described above for GC analysis. The monomer concentration was determined by comparison to the abundance peak of the standard and groups were compared by one-way ANOVA with Tukey's multiple test correction.
The optimum carbon concentration, temperature, and pH were determined for the PHA mcl producing bacteria. Naphthalene and anthracene were added to nutrient-limited media at different concentrations (200, 400, 600, 800, 1,400, and 1,600 mg/l) to evaluate the tolerance of the strains at 22°C for 5 d. The optical density (OD) of the strains was determined by UV spectrometry (Optizen 3220UV, Mecasys, Korea). The optimum temperature and pH of the strains were determined based on the OD observed after cultivating the organisms at seven different temperatures (5°C, 15°C, 20°C, 25°C, 30°C, 37°C, and 40°C) and five different pHs (5, 6, 7, 8, and 9) in nutrient broth media for 24 h.
3
Characterization of Hybrid Nanoparticles
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The chemical structure of the HACA and the degree of substitution of the CA were assessed by using 1H-NMR (Varian Unity 500MHz spectrometer, Palo Alto, CA, USA), in which CD3OD and D2O were used as the solvent. The hydrodynamic size and zeta potential of the HANPs and MPN-HANPs were observed by dynamic light scattering (DLS) using a Zetasizer Nano ZS90 (Malvern Ins. Worcestershire, UK) with a He-Ne 633nm laser at a 90° detection angle. The morphologies of the HANPs and MPN-HANPs were characterized by using HR-TEM (JEOL-2100F, Tokyo, Japan) operated at an accelerating voltage of 200 kV. For TEM images, the samples were dispersed in deionized water (DIW) and dropped onto a 300-mesh copper grid. The nanoparticles were then treated with 1% uranyl acetate for negative staining. The UV-visible (UV-Vis) absorption spectra were measured on a UV-vis spectrophotometer (Optizen 3220UV, Mecasys Co., Ltd., Daejeon, Korea). To investigate the stability of the nanoparticles, the changes in the scattering intensity of the HANPs and MPN-HANPs were assessed in the presence of sodium dodecyl sulfate (SDS) as a typical micelle-destabilizing agent. Briefly, the HANP and MPN-HANPs (1 mg/mL in DIW) were incubated with or without SDS (2.5 mg/mL). The resulting solutions were kept at room temperature and the scattering intensity of each solution was recorded using DLS as a function of time.
4
Fabrication of DOX-Loaded Hyaluronic Acid Nanoparticles
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The DOX-HANPs were prepared using an emulsion (water-in-oil) method. DOX·HCl (2 mg) was dissolved in dichloromethane, and three equivalents of triethylamine were slowly added while mixing. The DOX solution was added into an aqueous HANP (18 mg) solution to generate a water-in-oil emulsion. The emulsion was stirred overnight to evaporate the organic solvent in dark conditions. The solution was filtered using a 0.80 µm syringe filter to remove unloaded DOX aggregations. After filtration, the solution was dialyzed against DIW using membrane tubing (MWCO = 1 kDa), followed by lyophilization. The lyophilized DOX-HANPs (1.2 mg) were dispersed in 1 mL of DIW and the DOX was dissolved by adding 2 mL of DMF. After filtration of solution, the DOX amount of each of the samples was determined using the DOX standard curve. The amount of DOX loaded in the DOX-HANPs was determined using a UV-Vis spectrophotometer (Optizen 3220UV, Mecasys Co., Ltd., Daejeon, Korea) at a wavelength of 480 nm. To fabricate the DOX-MPN-HANPs, the MPN was coated on the DOX-HANPs in an identical manner to the MPN-HANPs. The hydrodynamic size and zeta potential of the DOX-MPN-HANPs were evaluated using a Zetasizer Nano ZS90, as described earlier.
5
Chlorophyll-a Extraction and Quantification
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To analyze chlorophyll-a, 5 mL aliquots obtained from each treatment were filtered through a 1.2-µm pore-sized GF/C filter (Whatman, Product No. 1823-047). The filters were frozen in the dark until chlorophyll-a was extracted and measured using spectrophotometry. The chlorophyll-a was extracted by placing the filter in 10 mL of 90% acetone for 24 h in the dark and was measured using a spectrophotometer (Model: Optizen 3220UV, Mecasys Co., Ltd., Daejeon, Korea) at the wavelengths of 630 nm, 645 nm, 663 nm, and 750 nm. Further, chlorophyll-a concentration was calculated using the following equation [49 ]:
6
Nanomaterial Characterization by TEM, UV-Vis, and Raman
7
Quantifying Gelatin Crosslinking Degree
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The degree of crosslinking was determined using 2,4,6-trinitrobenzensulfonic acid (TNBSA; Thermo Scientific, USA) by calculating the primary amine content of crosslinked and uncrosslinked gelatin samples. Briefly, 2–4 mg of gelatin was placed in a conical tube with 1.0 mL of 4% (w/v) sodium bicarbonate solution (NaHCO3, pH 8.5) and 1.0 mL of 0.5% (w/v) in methanol. After incubation at 40 °C for 2 h, the solution was treated with 3 mL of 6 M HCl, and the temperature was increased to 60 °C for 1.5 h to solubilize gelatin samples. The absorbance of the resulting solution was measured at 345 nm using a spectrophotometer (Optizen 3220UV; Mecasys Co., Ltd., Korea) after diluting with 4 volumes of deionized water. The degree of crosslinking was calculated using the following equation [9 (link)]:
where the subscripts c and u stand for the crosslinked and uncrosslinked gelatin, respectively (n = 3).
where the subscripts c and u stand for the crosslinked and uncrosslinked gelatin, respectively (n = 3).
8
Determination of Total Phenolic Content
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The total phenolic content of each extract was determined using the Folin-Denis method (25 ). Briefly, 0.5 mL of 1 N Folin-Ciocalteu reagent was added to 1 mL of extract mixed with 5 mL of distilled water and 1 mL of 95% ethanol. Then, 1 mL of 5% Na2CO3 solution was added after 5 min. The optical density (OD) at 725 nm was determined within 1 h using a UV-visible spectrophotometer (OPTIZEN 3220UV, Mecasys Co., Ltd., Daejeon, Korea). The phenolic content was determined using a standard curve of gallic acid.
9
Measurement of Bone Extract Turbidity
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A double beam spectrophotometer (Optizen 3220UV, Mecasys, Korea) was used for measuring turbidity. The bone extract sample was filtered using Whatman No. 1 filter paper. The turbidity of the filtered sample was measured at 590 nm; the blank was distilled water. Turbidity has been represented as % transmittance.
10
Comprehensive Blood Analysis Protocol
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Blood samples were collected from all animals under investigation by means of jugular vein puncture. Whole blood samples were analyzed for complete blood picture (Total white and red cells counts, hemoglobin, packed cell volume, mean corpuscular volume, mean corpuscular hemoglobin and mean corpuscular hemoglobin concentration) using Vet hematology analyzer (Medonic CA620–Sweden). Serum samples were analyzed for blood serum levels of glucose, total protein, sodium, chloride, potassium, urea and creatinine spectrophotometriclly (Optizen 3220 UV, Mecasys Co. Ltd, Korea) using diagnostic test kits (Spectrum Diagnostics, Cairo, Egypt and Spinreact, Spain) and according to the recommendation of the expert panel of the International Federation of Clinical Chemistry (IFCC).
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