All blood samples were taken in fasting condition at 9 a.m. at least 2 h after the last dose of ASA. Standard laboratory techniques were used for regular laboratory testing. Serum BDNF concentrations were measured using BDNF Quantikine Immunoassay (R&D Systems, USA) as previously described for our laboratory [14 (link)]. ELISA kits were also used to determine concentrations of the following parameters: serum TXB2 (EIA kits, Cayman Chemicals, Ann Arbor, MI, USA), von Willebrand factor (vWF) molecule (vWF: Ag), tumor necrosis factor (TNF)-α (Quantikine® HS ELISA Human TNF-α Immunoassay), interleukin (IL)-6 (Quantikine® HS ELISA Human IL-6 Immunoassay; both R&D Systems, Inc., Minneapolis, USA), soluble CD40 ligand (sCD40L; Human soluble CD40 Ligand Immunoassay, R&D Systems, Inc., NE, USA), and soluble P-selectin (human P-selectin/CD62P ELISA kit R&D Systems, Inc., Minneapolis, USA). High-sensitivity C-reactive protein (hsCRP) concentrations were assessed using a Cobas Integra 800 device (Roche, Basel, Switzerland), as previously described [21 (link),22 (link)]. The compliance with ASA treatment was defined according to previously described criteria [20 (link)].
Quantikine hs elisa human tnf α immunoassay
Manufactured by R&D Systems
Sourced in United States
The Quantikine® HS ELISA Human TNF-α Immunoassay is a solid-phase enzyme-linked immunosorbent assay designed to measure human tumor necrosis factor alpha (TNF-α) levels in cell culture supernates, serum, and plasma. The assay employs the quantitative sandwich enzyme immunoassay technique.
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Lab products found in correlation
4 protocols using quantikine hs elisa human tnf α immunoassay
1
Biomarker Measurement in Aspirin Therapy
2
Metabolic and Inflammatory Biomarkers
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Blood was drawn after an overnight fast; biochemical parameters were analyzed using standard laboratory methods as previously described [23 (link)]. Glycated hemoglobin A1C (HbA1c) levels were estimated as National Glycohemoglobin Standardization Program equivalent values (%), using the conversion formula established by the Japan Diabetes Society. Serum immune-reactive insulin (IRI) levels and serum C-peptide levels were measured by an electrochemiluminescence immunoassay (Roche Diagnostics K.K., Tokyo, Japan). High sensitivity TNFα (hsTNFα) and high sensitivity C-reactive proteins (hsCRP) were measured using a commercial ELISA kit (Quantikine HS ELISA human TNFα immunoassay and Quantikine ELISA human C-reactive protein immunoassay, R&D Systems, Inc., Minneapolis, MN, USA).
3
Cytokine Responses to Ultra-Marathon
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Before, immediately after and 90 minutes after completion of the ultra-marathon, blood samples were taken for determination of the plasma concentrations of IL-6, IL-10, IL-18, IL-1β and tumour necrosis factor alpha (TNF-α). The blood samples were collected in tubes containing EDTA. All samples were centrifuged within 15 minutes at 3000 rpm at 4°C, and stored at -70°C until further processed. High-sensitivity ELISA kits provided by R&D Systems (Minneapolis, MN, USA) were used for the pre-race blood samples for IL-6, IL-10 and TNF-α (Quantikine HS ELISA Human IL-6 Immunoassay, Quantikine HS ELISA Human IL-10 Immunoassay and Quantikine HS ELISA Human TNF-α Immunoassay, respectively) because these cytokines exist at very low levels in peripheral blood. Quantitative sandwich ELISA kits were used for the post-race and recovery blood samples for IL-6, IL-10, IL-18, IL-1β and TNF-α (R&D Systems Inc.; Minneapolis, MN, USA). Plasma concentration of IL-18 was determined using a Human IL-18 Instant ELISA kit (eBioscience, Bender MedSystems GmbH, Austria). All samples and provided standards were analysed in duplicate. Haematocrit was measured in duplicate. Whole blood samples (∼9 µl) were transferred to heparinized microcapillary tubes and analysed by an automated system following microcentrifugation. Haematocrit was used to calculate percent changes in plasma volume.
4
Platelet-rich plasma preparation for TEM
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Blood was collected with the use of a tourniquet into evacuated tubes with citrate or EDTA anticoagulants. Platelet-rich plasma (PRP) for transmission electron microscopy (TEM) was prepared by centrifugation of citrate-anticoagulated whole blood at 400–1600 g for 5 min or 200 g for 10 min (depending on the experimental protocol) with slow acceleration and no braking. Plasma TNF-α concentration was measured by enzyme-linked immunosorbent assay using a Quantikine™ HS ELISA human TNFα immunoassay (R&D Systems, Minneapolis, USA) according to the manufacturer’s instructions.
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