25 oh vitamin d total elisa kit

Blood samples were taken from the antecubital vein into the vacutainer tubes with silica clot activator. After centrifugation at 2000 × g for 10 min at 4°C, the serum was collected and stored in aliquots at -80°C until analysis.
Serum 25(OH)D₃ level was determined using the 25-OH Vitamin D total ELISA kit (DE1971, Demeditec Diagnostics, Germany) according to the manufacturer’s instructions. The intra-assay coefficients of variability (CVs) and inter-assay CVs reported by the manufacturer were 2–8% and 4–9%, respectively. Serum CRP, interleukin-6 (IL-6), tumor necrosis factor alpha (TNF-α), and interleukin-10 (IL-10) concentration were measured using the ELISA kit (DCRP00, HS600B, HSTA00E, and HS100C, respectively: R&D, United States) according to the manufacturer’s instructions. The intra-assay coefficients of variability (CVs) and inter-assay CVs reported by the manufacturer were 2–8% and 4–9%, respectively.

Blood samples were taken three times from the antecubital vein into the vacutainer tubes with a silica clot activator before, 15 min, and one hour after the tests. Additionally, in the WAnT group, the blood samples were taken 3 min after the first and second bout for lactate measurement. The samples were centrifuged at 2000g for 10 min at 4 °C. The separated serum and plasma samples were frozen and kept at − 80 °C until later analysis. The tubes containing the samples were number-coded to blind the laboratory personnel regarding the treatment group and the sequence of sample collection.
Serum 25(OH)D₃ concentration was determined by enzyme immunoassay method using 25-OH Vitamin D total ELISA kit (DE1971, Demeditec Diagnostics, Germany), according to the manufacturer’s instructions.
Plasma interleukin 6 (IL-6) concentration was determined by ELISA using a commercial kit (HS600, R&D Systems, USA).
Plasma parathyroid hormone (PTH) was determined using PTH intact ELISA kit (DE 3645, Demeditec Diagnostics, Germany).
Plasma non-esterified fatty acids and glycerol concentrations were determined by direct colorimetric methods using NEFA assay (FA115, Randox, United Kingdom) and Glycerol assay (GY105, Randox, United Kingdom).

Blood for biochemical research was collected four times: before supplementation (BS)—vitamin D3 or placebo, 5 weeks after supplementation 2 or 1 days before surgery ((AS), 4 weeks after surgery (BSVR), and 10 weeks after supervising rehabilitation (ASVR) in serum separation tubes (Becton Dickinson, Oxford, UK), The blood was collected at rest, while fasting, in the morning hours of 7:00–8:00 a.m. The samples for serum isolation were centrifuged at 2000× g and was stored at −80 °C until later analysis. Serum 25-OH Vitamin D3 was determined using the 25-OH Vitamin D total ELISA kit (DE1971, Demeditec Diagnostics, Kiel, Germany) according to the manufacturer’s instructions. The intra-assay coefficients of variability (CVs) and inter-assay CVs reported by the manufacturer were 2–8% and 4–9%, respectively.