Nf light advantage kit

Blood samples collected following decapitation were kept at room temperature for 30 min and then centrifuged for 10 min at 1500× g. The concentration of serum NfL was measured using the Simoa NfL assay (Quanterix, Quanterix Corp, Boston, MA, USA) NF-light Advantage kit (UmanDiagnostics, Umea, Sweden). The Simoa instrument uses bead-conjugated immunocomplex, which was applied to a multi-well array. The average number of enzymes per bead (AEB) was used for calibration by measurements on bovine NfL (UmanDiagnostics) serially diluted in assay diluent.

Mice were anaesthetized and blood samples were collected. Serum was frozen and stored at −80 °C until analysis. Sera were initially diluted 1:1000. When an obtained concentration was higher than 500 pg/mL, an additional dilution of 1:5000 was further tested. NFL concentrations were measured in duplicates by a single molecule array (Simoa) assay (Quanterix, Boston, MA, USA) and by a commercial kit (NF-light Advantage Kit, UmanDiagnostics Umea, Sweden), using a bead-conjugated immunocomplex. The immunocomplex was applied to a multi-well array designed to enable imaging of every single bead. The average number of enzymes per bead (AEB) of each sample was interpolated onto the calibrator curve constructed by AEB measurements on bovine NFL (UmanDiagnostics), serially diluted in an assay diluent. Samples were analyzed using one batch of reagents. Animal treatment information was blinded to the investigator performing the analysis.

Mice were anesthetized and blood samples were collected. The collected blood was incubated at room temperature for 30 min to enable clot formation and centrifuged at 1500 g for 10 min. The serum was frozen and stored at −80 °C until analysis. For NfL measurement, sera were initially diluted at 1:1000. When an obtained concentration was higher than 500 pg/mL, an additional dilution of 1:5000 was further tested. For tumor necrosis factor α (TNF-α), sera samples were diluted 1:8. NfL concentrations were measured in duplicates by a single molecule array (Simoa) assay (Quanterix, Boston, MA, USA) employing commercial kits (NF-light Advantage kit and mouse TNF-α discovery kit for HD-1/HD-X adjusted for SR-X, UmanDiagnostics Umea, Sweden), using a bead-conjugated immunocomplex. The immunocomplex was applied to a multi-well array designed to enable imaging of every single bead. The average number of enzymes per bead (AEB) of each sample was interpolated onto the calibrator curve constructed by AEB measurements on commercial NfL and TNF-α (UmanDiagnostics), serially diluted in an assay diluent. Samples were analyzed using one batch of reagents. Animal treatment information was blinded for the investigator performing the analysis.