Solutol® HS 15 (BASF, Ludwigshafen, Germany) is a mixture of free polyethylene glycol 660 and polyethylene glycol 660 hydroxystearate. Labrafac® WL 1349 (Gattefossé Group, Saint-Priest, France) is a mixture of capric and caprylic acid triglycerides. NaCl was purchased from Prolabo (Fontenay-sous-Bois, France). Lipoïd® S75-3 (Lipoïd GmbH, Ludwigshafen, Germany) is a soybean lecithin made of 69% phosphatidylcholine, 10% phosphatidylethanolamine, and other phospholipids. Milli-Q water was obtained from a Milli-Q-plus® system (Merck Millipore, Billerica, MA, USA). Chitosan oligosaccharide lactate 5,000 Da with a degree of deacetylation ≥75% was purchased from Sigma Aldrich (St Louis, MO, USA). The dialysis membrane was purchased from Spectrum Laboratories (Rancho Dominguez, USA) and had a molecular weight cut-off point equal to 50,000 Da. O-Phthalaldehyde (OPA) was purchased from Thermo Fisher Scientific (Waltham, MA, USA). MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) and MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assays (Cell Titer 96® kit) were purchased from Promega Corporation (Fitchburg, WI, USA).
O phthalaldehyde opa
Manufactured by Thermo Fisher Scientific
Sourced in United States
O-Phthalaldehyde (OPA) is a chemical compound used in analytical chemistry and biochemistry. It is a colorless, crystalline solid that is soluble in organic solvents. OPA is commonly used as a derivatizing agent for the detection and quantification of primary amines, including amino acids, by high-performance liquid chromatography (HPLC) or fluorescence spectroscopy.
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2 protocols using o phthalaldehyde opa
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Chitosan-Based Nanoparticle Formulation
2
Cell Viability and Protein Quantification
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Cell viability was tested after incubation with the different substances using PrestoBlue (Invitrogen, Carlsbad, CA, USA), a resazurin based cell viability reagent. Briefly, PrestoBlue was added 1∶10 to the cell culture medium at the end of the incubation time and incubated for additional 30 minutes at 37°C. The fluorescent signal was measured with a plate reader (HT Synergy, Biotek, Winooski, VT, USA) using an excitation of 530/25 nm and emission detection of 590/35 nm.
Total cellular protein was measured using o-phthalaldehyde (OPA), a primary amine-reactive fluorescent detection reagent from Thermo Scientific (Waltham, MA, USA). OPA (50 µl) was added to 50 µl sample and fluorescence was measured using a plate reader (HT Synergy, Biotek, Winooski, VT, USA) after 5 minutes incubation time using 360/40 nm for excitation and 460/40 nm for emission.
Total cellular protein was measured using o-phthalaldehyde (OPA), a primary amine-reactive fluorescent detection reagent from Thermo Scientific (Waltham, MA, USA). OPA (50 µl) was added to 50 µl sample and fluorescence was measured using a plate reader (HT Synergy, Biotek, Winooski, VT, USA) after 5 minutes incubation time using 360/40 nm for excitation and 460/40 nm for emission.
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