M-PER® Mammalian Protein Extraction Reagent (#78503 Pierce) was added to the isolated EV fraction using MagCapture Exosome Isolation Kit PS with Halt™ Protease and Phosphatase Inhibitor Cocktail (#78442 Thermo Fisher Scientific) and was mixed by vortexing for 15 min. The lysed EVs were filtered by 0.45 μm Spin-X centrifuge tube (#CLS8162 Corning). The EV t-tau and p-tau231 were measured using the Simoa Tau advantage kit (#101522 Quanterix, Lexington, KY, USA) and Simoa pTau-231 Advantage kit (#102292 Quanterix) on the Simoa HD-1 analyzer (Quanterix). All CSF-derived EV samples were diluted 10x with the Tau Calibrator Diluent (#101631 Quanterix) prior to the assays, to minimize matrix effects, and were analyzed in duplicate on one occasion. The relative concentration estimates of t-tau and p-tau231 were calculated according to the standard curve.
Simoa tau advantage kit
Manufactured by Quanterix
Sourced in United States
The Simoa Tau Advantage Kit is a diagnostic tool designed for the quantitative measurement of the tau protein in biological samples. It utilizes Simoa (Single Molecule Array) technology to provide highly sensitive and precise detection of tau levels. The kit is intended for research use only and does not make any claims about intended use or applications.
Automatically generated - may contain errors
Lab products found in correlation
Spin x centrifuge tube, by Corning (1 mentions)
Simoa ptau 231 advantage kit, by Quanterix (1 mentions)
Tau calibrator diluent, by Quanterix (1 mentions)
Simoa hd 1 analyzer, by Quanterix (1 mentions)
Vacutainer blood collection tube, by BD (1 mentions)
Simoa nf light advantage kit, by Quanterix (1 mentions)
Simoa hd 1, by Quanterix (1 mentions)
Single molecule array, by Quanterix (1 mentions)
Hd x instrument, by Quanterix (1 mentions)
3 protocols using simoa tau advantage kit
1
Quantification of Exosomal Tau Biomarkers
2
Quantification of Serum NfL and Tau
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All blood samples were collected from an antecubital vein, drawn into a standard gel 7 mL tube (BD Vacutainer blood collection tube, Becton Dickinson, Franklin Lakes, NJ, USA), before allowed to clot for 30 min. They were then centrifuged (3000 g for 10 min) before the serum was divided between two 1.5 mL Eppendorf tubes (Eppendorf, Hamburg, Germany).
The samples were then frozen within 2 h, after which one batch was used for the analyses in the original study (19) (link) and the other batch was stored at -80°C for later analyses.
During the summer of 2018, the samples were thawed to 20°C and centrifuged (20 000 g for 10 min). Serum NfL and serum tau concentrations were then measured using ultrasensitive single molecule array technology (SIMOA-HD1, Quanterix Corporation, Lexington, MA, USA). Simoa™ NF-Light Advantage Kit and Simoa™ Tau Advantage Kit (Quanterix Corporation, Lexington, MA, USA) were used for NfL and tau protein quantification, respectively. Each measurement was done in duplicate, with a limit of detection of 38 fg/mL for serum NfL and 20 fg/mL for serum tau.
All analyses were performed blinded at the Institute of Neuroimmunology, Slovak Academy of Sciences, Bratislava, Slovakia, and in accordance with the manufacturer's recommendations.
The samples were then frozen within 2 h, after which one batch was used for the analyses in the original study (19) (link) and the other batch was stored at -80°C for later analyses.
During the summer of 2018, the samples were thawed to 20°C and centrifuged (20 000 g for 10 min). Serum NfL and serum tau concentrations were then measured using ultrasensitive single molecule array technology (SIMOA-HD1, Quanterix Corporation, Lexington, MA, USA). Simoa™ NF-Light Advantage Kit and Simoa™ Tau Advantage Kit (Quanterix Corporation, Lexington, MA, USA) were used for NfL and tau protein quantification, respectively. Each measurement was done in duplicate, with a limit of detection of 38 fg/mL for serum NfL and 20 fg/mL for serum tau.
All analyses were performed blinded at the Institute of Neuroimmunology, Slovak Academy of Sciences, Bratislava, Slovakia, and in accordance with the manufacturer's recommendations.
3
Measuring Neurological Biomarkers in CSF
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Venous blood samples were obtained at empirically determined regular time points up to about 48 h after intrathecal administration of gadobutrol at the time of the MRI acquisitions and stored in the refrigerator (4 oC) for a few hours before centrifuge and thereafter storage in an ultra-freezer (−80 oC). Aβ40, Aβ42, GFAP, and NfL concentrations were measured using the Single molecule array (Simoa) Human Neurology 4-Plex E (N4PE) assay, whilst T-tau was measured using the Simoa Tau Advantage kit according to instructions from the kit manufacturer (Quanterix, Billerica MA). All measurements were performed on an HD-X instrument (Quanterix, Billerica MA) in one round of experiments using one batch of reagents by board-certified laboratory technicians who were blinded to the clinical data. Intra-assay coefficients of variation were below 10%.
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