Uv vis spectrophotometry

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UV-Vis spectrophotometry is an analytical technique that measures the absorption of ultraviolet (UV) and visible (Vis) light by a sample. It is used to determine the concentration of a substance in a solution by measuring the amount of light absorbed at a specific wavelength.

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Lab products found in correlation

Alexa fluor 488 nhs, by Thermo Fisher Scientific (1 mentions) 20k mwco dialysis cups, by Thermo Fisher Scientific (1 mentions) Cary 100 series, by Agilent Technologies (1 mentions) Primescript 1st strand cdna synthesis kit, by Takara Bio (1 mentions) Tri regent, by Merck Group (1 mentions) C myc, by Bioneer (1 mentions)

7 protocols using uv vis spectrophotometry

Quantifying Quercetin Encapsulation in Liposomes

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Standard curves were prepared using stock solution of 1 mM QU and dimethyl sulfoxide (DMSO) as a blank. The calibration curves were linear (R² > 0.996) against the QU concentration (1–10 µg mL⁻¹).
Entrapment efficiency of QU-loaded liposome was determined by assessing the difference between the total amount of QU and the amount of free QU present in the liposome. The amount of QU encapsulated in the QU-ML-based liposomes was identified by an indirect method via centrifugation technique using a centrifuge (Model: 3740, KUBOTA MFG. CORP, Tokyo, Japan) at 5000 rpm for 15 min at 4 °C. The experiment of QU quantification was carried out at 370 nm [27 (link)]. The concentration of QU was measured via UV/V is spectrophotometry (Agilent Technologies, Basel, Switzerland) at 370 nm. The EE_QU was then calculated according to Equation (2):

% E E_{Q U} = \frac{(W_{t o t a l} - W_{f r e e}) \times 100}{W_{t o t a l}}

where W_total is the total QU weight in liposomes suspension; W_free is the weight of free QU.

Toopkanloo S.P., Tan T.B., Abas F., Alharthi F.A., Nehdi I.A, & Tan C.P. (2020). Impact of Quercetin Encapsulation with Added Phytosterols on Bilayer Membrane and Photothermal-Alteration of Novel Mixed Soy Lecithin-Based Liposome. Nanomaterials, 10(12), 2432.

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Affibody-Enzymes Fluorescent Labeling

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The affibody-enzymes (N23BP-CodA and WT-CodA) were conjugated to 1:10 molar excess NHS-AlexaFluor 488 (Thermo Fisher) and the pH-sensitive NHS-pHAb (Promega) in separate tubes for 2 h at RT in the dark in PBS. The mixtures were then transferred to a 20k MWCO dialysis cups (Thermo Fisher) and excess dye was removed by dialysis. After 36 h dialysis, the degree of labeling for both the dye-conjugated affibody fusions were calculated using UV-Vis spectrophotometry (Agilent Technologies, Cary Series 100), by using the absorbance at 545 nm for the endotracker pHAb and 495 nm for AF488 as compared to the absorbance at 280 nm for both affibody-fusions.

Heerklotz D., Döring P., Bonzelius F., Winkelhaus S, & Nover L. (2001). The Balance of Nuclear Import and Export Determines the Intracellular Distribution and Function of Tomato Heat Stress Transcription Factor HsfA2. Molecular and Cellular Biology, 21(5), 1759-1768.

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Antioxidant Activity of PSEO

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The inhibition of β-carotene bleaching by PSEO was assessed as described by Marco (1968) . First, a β-carotene/linoleic acid emulsion was prepared by dissolving 0.5 mg of β-carotene, 25 μL of linoleic acid and 200 μL of Tween 40 in 1 mL of chloroform. The chloroform was then totally evaporated under vacuum using a rotatory evaporator at 50 °C. A volume of 100 mL of distilled water was added and the resulting emulsion was vigorously stirred. Thereafter, 2.5 mL of the β-carotene/linoleic acid emulsion were mixed with 0.5 mL of PSEO at different concentrations (100-700 µg/mL). The absorbance was measured at 470 nm before and after incubation at 50 °C for 2 h by UV-Vis spectrophotometry (Agilent, USA). BHA was the positive control. Tests were conducted in duplicate and the β-carotene bleaching inhibition was calculated using the following formula:
where OD 0 and OD t were the absorbances of the test sample (PSEO) measured before and after incubation, respectively; and OD' 0 and OD' t were the absorbances of the control measured before and after incubation.

Ksouda G., Sellimi S., Merlier F., Falcimaigne-Cordin A., Thomasset B., Nasri M, & Hajji M. (2019). Composition, antibacterial and antioxidant activities of Pimpinella saxifraga essential oil and application to cheese preservation as coating additive. Food chemistry, 288.

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Astaxanthin Modulates Stemness and Proliferation

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Post 24 hours exposure of ADMSCs with 0 and 0.5 ng/ml of astaxanthin, the total RNA was isolated using TRI regent (Sigma-Aldrich, MO, IL, USA). The total RNA concentration was measured using UV/Vis-spectrophotometry (BioTek, Winooski, VT, USA) at 260 nm. cDNA was synthesized from the RNA using the reverse transcription method PrimeScript 1^st strand cDNA Synthesis Kit (Takara, Shiga, Japan). Template DNA was then used in gene-specific PCR, wherein synthesized cDNA using oligo-dT primer was amplified by 40 cycles (initial denaturation, denaturation, annealing, and extension: 98 °C, 1 min, 98 °C, 10 sec; 55–60 °C, 30 sec; 72 °C, 1 min). The expression of stemness-related genes (SOX2 and KLF4 (Bioneer, Alameda, California, USA)) and proliferation-related genes (Rex1, c-MYC, and Wnt3a, (Bioneer, Alameda, California, USA)) were studied, wherein Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was utilized as a housekeeping gene. Details of the primers are listed in Table 1. Further, aliquots of PCR product were electrophoresed on 1.5% agarose gels, PCR fragments were stained by loading STAR dye (Dynebio, Gyeonggi-do, South Korea) and detected by the gel documentation system (Daihan Scientific, Seoul, South Korea). All gene expression experiments were performed in triplicates.

Choi B.Y., Chalisserry E.P., Kim M.H., Kang H.W., Choi I.W, & Nam S.Y. (2019). The Influence of Astaxanthin on the Proliferation of Adipose-derived Mesenchymal Stem Cells in Gelatin-Methacryloyl (GelMA) Hydrogels. Materials, 12(15), 2416.

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Evaluating HA-Ge Mineralization in SBF

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The efficiency of HA-Ge in mineralization was evaluated by the method of Kokubo et al. using simulated body fluid (SBF) [39 (link)]. The SBF (pH 7.4) was prepared exactly following the same steps and recipes, as recommended by Kokubo et al. [39 (link)]. The samples were incubated vertically in 50 mL falcon tubes with SBF for 28 days at 37 °C with changing the medium every 3 days once. Then, the samples were gently washed with distilled water three times and stained with an alizarin red stain to visualize the mineral deposition on the surface. The amount of staining rate was optically measured by UV-Vis spectrophotometry (Bio Tek Instruments Inc., Winooski, VT, USA) at 450 nm by subtracting the control value (samples without SBF treatment). For quantification, the alizarin red stain was dissolved in a mixture of methanol (20%) and acetic acid (10%) in water for 20 min [40 (link)].

Elango J., Bushin R., Lijnev A., De Aza P.N., Martínez C.P., Marín J.M., Hernandez A.B., Olmo L.R, & Val J.E. (2022). The Effect of Germanium-Loaded Hydroxyapatite Biomaterials on Bone Marrow Mesenchymal Stem Cells Growth. Cells, 11(19), 2993.

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Characterization of MPEO Nanoparticles

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To calculate the EE % and LC%, rst the maximum absorbance wavelength of MPEO was obtained by UV-Vis spectrophotometry (Bio Tek, USA) in a wavelength range between 200 to 600 nm. For this purpose, the diluted concentration of MPEO in methanol was analyzed in an absorbance range of 1, and methanol was used as a blank. Then, 1 ml of the MPEO-N nanoparticle suspension (equivalent to 1 g of formulation) was dissolved in 1 ml of isopropanol, and the amount of MPEO in the solvent was determined by a UV-Vis spectrophotometer at the wavelength of maximum absorbance of MPEO (using isopropyl as blank and corresponding calibration curve). Finally, the LC and EE values of MPEO-N nanoparticles were calculated based on the following equations [11] : × 100

Hamzian N., Nickfarjam A., Shams A., Haghiralsadat F, & Najmi-Nezhad M. (2024). Radioprotective effect of nanoniosome loaded by Mentha Pulegium essential oil on human peripheral blood mononuclear cells exposed to ionizing radiation. Drug development and industrial pharmacy, 50(3).

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Conjugation of Protein A and scFv-Fc Antibodies to Nanoparticles

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Protein A and scFv-Fc antibodies were conjugated to Nano Act™ CNBs (Asahi Kasei, Japan) using a CNB conjugation kit (DCN Diagnostics, CA, USA). Briefly, protein A and antibodies (0.5 mg/ml in 120 μl conjugation buffer) were mixed with CNB (0.5% CNB in 120 μl conjugation buffer) and incubated for 2 h at 37 °C. Then, 7.2 ml blocking buffer were added to block the CNB surface. After blocking for 2 h at 37 °C, unconjugated CNB was removed and the protein A and antibody-CNB conjugants were washed with 7.5 ml wash buffer by centrifugation at 14,400 g for 20 min at 4 °C. Finally, the pellets were suspended gently in 300 μl wash buffer. The concentration of the protein A and antibody-CNB conjugants was measured using UV–vis spectrophotometry (BioTek Instruments, Inc., VT, USA) at an absorption intensity of 554 nm.

Kim H.Y., Lee J.H., Kim M.J., Park S.C., Choi M., Lee W., Ku K.B., Kim B.T., Changkyun Park E., Kim H.G, & Kim S.I. (2020). Development of a SARS-CoV-2-specific biosensor for antigen detection using scFv-Fc fusion proteins. Biosensors & Bioelectronics, 175, 112868.

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