Saliva was collected twice, by passive drool method, in small polypropylene tubes. Samples were immediately stored at -18°C, before further analyses. Enzyme-linked immunosorbent assay kits (Expanded Range High Sensitivity Salivary Cortisol Enzyme Immunoassay Kit No. 1–3002, Salimetrics, UK) were used to determine salivary cortisol levels. A 96-well microplate spectrophotometer (Multiska FC Microplate Photometer, Thermo Fisher, Germany) was used to measure absorbance at 450nm. Each well, containing samples or cortisol standards and controls, was duplicated. Intra-assay precision was determined from the mean of replicates with a single microplate (coefficient of variation 4.8 ±1.2%). Inter-assay precision was determined from the mean of average duplicates based on the cortisol standards wells for calibration, provided in the ELISA kit, used in separate runs (coefficient of variation 5.4 ±3.6%), but prepared from the same stock solution and dilutions. The concentrations of unknown samples, expressed in nmol/L, were computed by interpolation using a 4-parameter non-linear regression curve fit, as recommended by Salimetrics’ protocol.
Multiska fc microplate photometer
Manufactured by Thermo Fisher Scientific
Sourced in Germany
The Multiska FC Microplate Photometer is a compact and versatile spectrophotometer designed for microplate-based absorbance measurements. It can perform measurements across a wide range of wavelengths to support various assays and applications.
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Lab products found in correlation
2 protocols using multiska fc microplate photometer
1
Salivary Cortisol Quantification: Precision Assay Approach
2
SARS-CoV-2 Spike Protein ELISA
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The wells of a 96-well plate pre-coated with 100 ng of S protein of SARS-CoV-2, SARS-CoV (Sino Biological Inc., Catalog Number: 40634-V08B), or MERS-CoV (Sino Biological Inc., Catalog Number: 40069-V08B) were blocked with 0.25% gelatin in PBS buffer containing 0.05% Tween-20 (PBST), and then incubated with 100 μL of indicated mAbs at room temperature for 1 h. After washing of the 96-well plate three times with PBST, the HRP-conjugated goat anti-mouse IgG (H + L) (KPL) was added to each well and incubated at room temperature for 1 h. After washing of the 96-well plate three times with PBST, 75 μL of 3,3′,5,5′-tetramethylbenzidine substrate (BD Bioscience) was added to each well for signal detection, followed by addition of 75 μL of 2 N H2SO4 to terminate the reactions. Results are evaluated quantitatively by measuring the absorbance at 450 nm by the Multiska FC Microplate Photometer (Thermo Fisher Scientific).
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