All CSF samples were collected by lumbar puncture in the L3/L4 or the L4/L5 inter-space, in the morning. The first 12 mL of CSF were collected in a polypropylene tube and immediately transported to the local laboratory and centrifuged (10 min at 1800g at + 20 °C). The supernatant was gently mixed to avoid possible gradient effects, aliquoted in polypropylene tubes, and stored at − 80 °C. CSF t-tau and p-tau concentrations were measured using ELISA (INNOTEST® htau Ag and PHOSPHO_TAU (181P), Fujirebio) as previously described [13 (link), 43 (link)]. CSF Aβ1-42 was measured using an ELISA (INNOTEST® Aβ1-42), specifically constructed to measure Aβ starting at amino acid 1 and ending at amino acid 42 [6 (link)].
Phospho tau 181p
Manufactured by Fujirebio
Sourced in Belgium
PHOSPHO-TAU[181P] is a laboratory assay that measures the concentration of phosphorylated tau protein at the 181st amino acid residue. This analyte is used as a biomarker in research applications.
Automatically generated - may contain errors
Lab products found in correlation
Innotest β amyloid1 42, by Fujirebio (6 mentions)
Htau ag, by Fujirebio (6 mentions)
β amyloid1 42, by Fujirebio (5 mentions)
Innotest htau ag, by Fujirebio (3 mentions)
Multiskan mk3, by Thermo Fisher Scientific (3 mentions)
Enzyme linked immunosorbent assay, by Fujirebio (2 mentions)
Innotest amyloid β42, by Fujirebio (2 mentions)
V plex aβ peptide panel 1 6e10 kit, by Mesoscale (1 mentions)
Kaspar pcr snp genotyping system, by LGC (1 mentions)
Enzyme linked immunosorbent assay, by UmanDiagnostics (1 mentions)
Aβ1 42, by Fujirebio (1 mentions)
Innotest assays, by Fujirebio (1 mentions)
Lumipulse g600ii instrument, by Fujirebio (1 mentions)
Pcr 02 c, by Corning (1 mentions)
Polypropylene tube, by Sarstedt (1 mentions)
Nf light elisa kit, by UmanDiagnostics (1 mentions)
Innobia alzbio3 immunoassay, by Fujirebio (1 mentions)
Msd human aβ peptide ultra sensitive kit, by Mesoscale (1 mentions)
28 protocols using phospho tau 181p
1
CSF Biomarker Quantification Protocol
2
CSF Protein Biomarker Measurement Protocols
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CSF t-tau and p-tau (tau phosphorylated at threonine 181) concentrations were measured with sandwich enzyme-linked immunosorbent assays (ELISAs) (INNOTEST htau Ag and PHOSPHO_TAU [181P], Fujirebio [formerly Innogenetics], Ghent, Belgium) (22 (link),23 (link)). CSF Aβ42 was measured with a sandwich ELISA (INNOTEST Aβ1–42) specifically constructed to measure Aβ starting at amino acid 1 and ending at amino acid 42 (24 (link)). For NfL, an in-house sandwich ELISA with capture and detection antibodies that were directed against the central rod domain of the protein (NfL 21 and NfL 23, respectively) was used (25 (link)). An in-house ELISA method (26 (link)) was used to measure CSF Ng.
3
Cerebrospinal Fluid Biomarkers Analysis
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Cerebrospinal fluid (CSF) samples were collected in the morning through LP in L3/L4 or L4/L5 interspaces. A total of 10 ml of CSF was collected in a polypropylene tube and immediately transported to the laboratory and centrifuged at 1 800g at 20° C. The supernatant was gently mixed to avoid possible gradient effects, aliquoted in polypropylene tubes, and stored at −70° C. CSF phosphorylated tau at threonine 181 (p-tau) concentrations were determined using sandwich enzyme-linked immunosorbent assay (INNOTEST htau Ag and PHOSPHO_TAU (181P; Fujirebio, Ghent, Belgium) (40 (link)). We used the CSF Aβ42/40 ratio as a biomarker of amyloid-beta pathology. The CSF Aβ42/40 ratio was obtained using the V-PLEX Aβ peptide Panel 1 (6E10) kit (Meso Scale Discovery, Rockville, MD) (41 (link)). This variable was treated continuously in the main analyses. To characterize the sample, following Samuelsson et al. (42 (link)) we classified CSF p-tau and Aβ42/40 values into positive and negative using the following cutoff values: ≥80 pg/mL for p-tau and ≤0.082 for Aβ42/40. The APOE-ε4 allele was determined using the KASPar PCR SNP genotyping system (LGC Genomics, Hoddesdon, Herts, UK) as described by Skoog et al. (43 (link)) To characterize the sample, participants were classified as APOE-ε4 carriers if they had at least 1 ε4 allele.
4
Cerebrospinal Fluid Biomarker Analysis
5
Standardized Biomarker Sampling and Analysis
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Fasting CSF samples for the Wisconsin ADRC cohorts and WRAP were collected via lumbar puncture13 following the same protocol and by the same group of well‐trained individuals.13 Samples were sent together in two batches to the lab of Drs. Blennow and Zetterberg in Sweden, where commercially available enzyme‐linked immunosorbent assay (ELISA) methods were used to quantify CSF t‐tau, p‐tau, and amyloid beta 1‐42 (Aβ42; INNOTEST assays HTAU AG, PHOSPHO‐TAU[181P], and Aβ1‐42, respectively; Fujirebio).13 The batch‐adjusted predicted values for CSF biomarkers were used for all analyses.17 In WRAP, fasting blood samples were collected in ethylenediaminetetraacetic acid (EDTA) tubes; the plasma was pipetted off within 1 hour of collection and stored at –80°C.12 A total of 141 longitudinal samples from 123 individuals in WRAP with plasma metabolites were available for the main analysis. In the Wisconsin ADRC, blood samples were collected in heparin tubes, which could influence metabolite values; as such, plasma metabolomics data have not been generated in Wisconsin ADRC blood samples. Further details of how plasma and CSF samples were processed are explained in an earlier study.12
6
Cerebrospinal Fluid Biomarker Measurement
7
Circadian Rhythm-Adjusted CSF Biomarkers
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All patients underwent lumbar puncture between 8:00 and 10:00 am to avoid variations related to the circadian rhythm. Samples were collected in polypropylene tubes, centrifuged at 2000 × g for 10 min at 4°C and stored at −80°C until use. The levels of CSF Aβ42 (Innotest® β-Amyloid (1-42)), t-tau (Innotest® hTAU Ag) and p-tau (Innotest® Phospho-Tau (181P)) were determined by the enzyme immunoassay method according to the manufacturer’s instructions (Fujirebio Europe, Ghent, Belgium). All samples were measured in duplicate and expressed in pg/ml. Samples were obtained with support from IRBLleida Biobank (B.0000682) and PLATAFORMA BIOBANCOS PT17/0015/0027.
8
CSF Biomarker Quantification Protocol
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CSF collection and processing methods are described elsewhere (Niemantsverdriet et al., 2017 (link)). CSF concentrations of Aβ1–42, Aβ1–40, T-tau, and P-tau181 were determined with commercially available single-analyte ELISA (INNOTEST® β-AMYLOID(1–42), β-AMYLOID(1–40), hTAU-Ag, and PHOSPHO-TAU(181P), respectively; Fujirebio Europe) as routinely performed in the BIODEM lab (Somers et al., 2016 (link)). The laboratory technician performing the biomarker analyses was blinded to clinical diagnosis. In addition, the Aβ1–42/Aβ1–40, Aβ1–42/T-tau, and Aβ1–42/P-tau181 ratios were calculated. CSF biomarker results were not included in the consensus clinical diagnosis made by the panel.
9
CSF Biomarker Quantification Protocol
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CSF was obtained by lumbar puncture between L3/4, L4/5, or L5/S1 intervertebral space, using a 25-gauge needle and a syringe [36 ]. The samples were collected in polypropylene microtubes and centrifuged at 1800g for 10 min at 4 °C. Thereafter, the samples were frozen at − 20 °C until manual analyses of Ab42, tau, and p-tau were performed using sandwich ELISAs [Innotest assays: β-amyloid1-42, tTAU-Ag, and PhosphoTAU-181p; Fujirebio (formerly Innogenetics)] at the Neurochemistry Laboratory of the Department of Clinical Chemistry of VUmc. As the median CSF Aβ42 values of our cohort have been gradually increasing over the years [37 (link)], we determined CSF amyloid-β status using Aβ42 values that had been adjusted for the longitudinal upward drift. We used a uniform cut-off of 813 pg/mL to dichotomize CSF data [38 (link)].
10
Biomarker Quantification in ADRC Cohorts
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Fasting CSF samples for the Wisconsin ADRC cohorts and WRAP were collected via lumbar puncture [13] (link) following the same protocol and by the same group of well-trained individuals [13] (link). Samples were sent together in two batches to the lab of Drs.
Blennow and Zetterberg in Sweden, where commercially available enzyme-linked immunosorbent assay (ELISA) methods were used to quantify CSF t-tau, p-tau, and amyloid-beta 1-42 (Aβ42) (INNOTEST® assays HTAU AG, PHOSPHO-TAU[181P], and β-amyloid1-42, respectively; Fujirebio, Ghent, Belgium) [13] (link). The batch-adjusted predicted values for CSF biomarkers were used for all analyses [17] (link).
In WRAP, fasting blood samples were collected in ethylenediaminetetraacetic acid (EDTA) tubes; the plasma was pipetted off within one hour of collection and stored at -80°C [12] (link).
A total of 141 longitudinal samples from 123 individuals in WRAP with plasma metabolites were available for the main analysis. In (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
the Wisconsin ADRC, blood samples were collected in heparin tubes, which could influence metabolite values; as such, plasma metabolomics data have not been generated in Wisconsin ADRC blood samples. Further details of how plasma and CSF samples were processed are explained in an earlier study [12] (link).
Blennow and Zetterberg in Sweden, where commercially available enzyme-linked immunosorbent assay (ELISA) methods were used to quantify CSF t-tau, p-tau, and amyloid-beta 1-42 (Aβ42) (INNOTEST® assays HTAU AG, PHOSPHO-TAU[181P], and β-amyloid1-42, respectively; Fujirebio, Ghent, Belgium) [13] (link). The batch-adjusted predicted values for CSF biomarkers were used for all analyses [17] (link).
In WRAP, fasting blood samples were collected in ethylenediaminetetraacetic acid (EDTA) tubes; the plasma was pipetted off within one hour of collection and stored at -80°C [12] (link).
A total of 141 longitudinal samples from 123 individuals in WRAP with plasma metabolites were available for the main analysis. In (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
the Wisconsin ADRC, blood samples were collected in heparin tubes, which could influence metabolite values; as such, plasma metabolomics data have not been generated in Wisconsin ADRC blood samples. Further details of how plasma and CSF samples were processed are explained in an earlier study [12] (link).
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