NO has an extremely short half-life and quickly metabolizes to nitrate and nitrite. Therefore, we utilized a colorimetric nitrite assay kit (Sigma–Aldrich) to measure nitrite levels, which was used to indicate NO production in a more stable format. All nitrite assay experiments were performed per the manufacturer’s protocol. In brief, an equal volume of detection reagents were added to 96-well plates with Hank’s balanced salt solution (in mM: 120 NaCl, 5 KCl, 1 MgCl2, 5.5 glucose, 10 HEPES, 1.23 CaCl2, pH 7.4) containing CMs. Azo dye production was quantified by absorbance at 540 nm in room temperature using an Emax precision microplate reader (Molecular Devices). Nitrate production was quantified utilizing standardized reagents which produced a standard curve against which we plotted our absorbance recording. Results were normalized to each other by lysing the individual samples and subsequential protein concentration assessment. Results are reported as the amount of nitrite produced per mg of sample protein.
Emax precision
Manufactured by Molecular Devices
Sourced in United States
The Emax Precision is a high-performance microplate reader designed for accurate and precise absorbance measurements. It offers a wide range of wavelengths and supports multiple detection modes, enabling diverse applications in scientific research and analysis.
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Lab products found in correlation
6 protocols using emax precision
1
Colorimetric Nitrite Assay for NO Quantification
2
Glutamate-Induced Cell Viability Assay
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Cell viability in HT-22 cells was investigated according to a previously reported method [10 (link)]. Briefly, the HT-22 cells were seeded into 96-well plates at a density of 3 × 103 cells/well for 24 h, with each well containing 100 μL of medium. The cells were treated with glutamate (5 mM) in the presence or absence of RDP (5–50 μg/mL). After 11 h of incubation, cell viability was evaluated using the EZ-Cytox Cell Viability Assay Kit following the manufacturer’s instructions. The absorbance was measured by an Emax Precision microplate reader (Molecular Devices, Sunnyvale, CA, USA) at 450 nm. The percentage of surviving cells was calculated relative to the values of vehicle control group.
3
Glutamate-Induced Cytotoxicity Assay
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Cell viability was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay kits, according to the manufacturer’s instructions MTT assay kit, USB Corporation, Cleveland, OH, USA). HT-22 cells were seeded into 48-well plates at a density of 1×104 cells per well for 24 h, with each well containing 500 μL cell culture medium. Cells were treated with glutamate (5 mM) in the presence or absence of WSP (10∼100 μg/mL). After 11 h incubation, MTT solution (0.2 mg/mL) was added to each well, and the plates were incubated at 37°C for 30 min. Then, the absorbance was read using an Emax Precision microplate reader (Molecular Devices, Sunnyvale, CA, USA) at 550 nm. The percentages of surviving cells were determined relative to control cells.
4
LDH Leakage Assay for Cell Viability
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LDH leakage was assessed using a LDH assay kit, according to the manufacturer’s instructions (Roche Diagnostics, Indianapolis, IN, USA). Briefly, HT-22 cells were preincubated with or without WSP (0∼100 μg/mL) for 30 min in 48-well plates before challenge with glutamate. Then, the medium of each well was removed and placed into 96-well plates, and the kit reagents was added and left to react for 10 min. The absorbance was then read using an Emax Precision microplate reader (Molecular Devices) at 490 nm and the percentage of surviving cells determined relative to control cells.
5
Sialyllactose Effects on THP-1 Cell Viability
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THP-1 cells were seeded into 96-well plates and treated with α2,3- and α2,6-sialyllactose (0.1 μM, 1 μM, 10 μM, 100 μM, and 300 μM) for 24 h. Cell viability was measured by adding 10 μl of D-Plus CCK (cell viability, proliferation, and cell cytotoxicity assay kit) reagent (Dongin LS, Seoul, Republic of Korea). The cells were incubated for 1 h at 37°C, and the optical density (OD) at 450 nm was determined using an EMax precision microplate reader (Molecular Devices, Sunnyvale, CA, USA).
6
Serum Insulin Quantification in Mice
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