The blood was collected immediately following decapitation. The collected blood was incubated in room temperature for 30 min and centrifuged (1500 g, 10 min, 25 °C). The serum was collected and stored at – 80 °C until the assay. Mouse serum NfL concentration was determined using the Simoa NfL assay as previously described [32 (link)] (Quanterix, Quanterix Corp, Boston, MA, USA) (UD2, UmanDiagnostics) in a Simoa instrument (Quanterix) 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 samples was interpolated onto the calibrator curve constructed by AEB measurements on bovine NfL (UmanDiagnostics) serially diluted in assay diluent. Samples were analyzed using one batch of reagents and animal treatment information was blinded to the one performing the analysis.
Bovine nfl
Manufactured by UmanDiagnostics
Bovine NfL is a laboratory diagnostic tool used to detect and measure the levels of neurofilament light (NfL) protein in bovine samples. NfL is a biomarker for neurological health and can provide insights into various neurological conditions in cattle.
Automatically generated - may contain errors
3 protocols using bovine nfl
1
Serum NfL Quantification in Mice
2
Serum NfL Quantification via Simoa Assay
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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.
3
Serum NFL Concentration Measurement
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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.
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