A surrogate neutralization assay was used because it does not require the use of active virus and was validated extensively using patient samples – showing high sensitivity and specificity for SARS-CoV-2 spike protein.51 (link) Abs were prepared by serial dilution in assay buffer (3% bovine serum albumin (BSA) + PBS). Samples were incubated with 0.1 mg/mL of biotinylated spike glycoprotein at 1:1 (v/v) for 1 hr with shaking. After incubation, 25 µL of the mixture was added to Streptavidin Gold multi-array 96-well plates (Meso Scale Discovery®) and incubated for 1 hr with shaking. After incubation, 25 mL of ruthenium labeled ACE2 at 2 mg/mL were added and incubated for 1 hr with shaking. Plates were washed with PBS prior to addition of 150 mL of 2 X MSD read buffer to read the plate on Mesoscale Sector S 600 reader. MSD data was analyzed and IC50 values were calculated using Graphpad Prism software.
Sector s 600 reader
Manufactured by Mesoscale
The Sector S 600 reader is a laboratory instrument used for detecting and quantifying various analytes in samples. It functions by measuring the intensity of light emitted from labeled molecules within the samples. The core purpose of the Sector S 600 reader is to provide accurate and reliable data for research and diagnostic applications.
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Lab products found in correlation
3 protocols using sector s 600 reader
1
Surrogate SARS-CoV-2 Neutralization Assay
2
Measuring Spinal Cord IL-6 Levels
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Thoracic and lumbar spinal cords from treated mice were snap frozen in liquid nitrogen and pulverized using hammer and pistil. Pulverized spinal cord was weighed and 1 ml/100 mg MSD Tris Lysis Buffer (R60TX-3) was added according to the manufacturers’ protocol. Samples were passed six times through a 25-gauge needle and kept on ice for 25 min prior to centrifugation at 14,000 rpm for 10 min at 4 °C. Supernatant was immediately snap frozen in liquid nitrogen and analyzed with an commercial ELISA system (U-PLEX Mouse IL-6, manufacturer #B22TX-2) according to the manfacturers instuctions on a Sector S 600 reader (All: Meso Scale, Rockville, Maryland).
3
Quantifying Amyloid-Beta Levels
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The levels of Aβ40 and Aβ42 were determined by electrochemiluminescence (ECL) assay using 96-well MULTI-ARRAY Multiplex Kits (Cat# K15199E-2, Meso Scale Diagnostics, Gaithersburg, MD). The TBS- and guanidine-soluble fractions of brain lysates were processed according to the manufacturer’s instructions. Aβ levels were measured using a SECTOR S 600 reader (Meso Scale Diagnostics).
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