Human free bdnf elisa kit

We measured serum and not plasma BDNF concentrations as serum measurements have been used more frequently than plasma measurements in previous studies, which improves comparability of our results (serum concentrations combine blood-borne bound and unbound BDNF, and plasma concentrations display only the unbound proportion; Dinoff et al., 2017 (link)). BDNF was measured in participants with at least four of six successful visits in serum by the human free BDNF ELISA kit (R&D Systems, Minneapolis, MN, United States) according to the instructions of the manufacturer with a dilution factor of 1:3 (standard and sample). All standards, samples, and controls were run once. A sigmoid curve was fitted on the resulting data, and the sample concentrations were calculated by the Dynex DS2 software.
Platelet counts were measured with Sysmex fluorescence flow cytometry (XE-2100D, Sysmex Xtra 2/2008) according to the instructions of the manufacturer and in line with the ICSH reference methods (International Council for Standardization in Hematology).
In a second step, results of serum BDNF concentrations and platelet count were corrected according to the method of Dill and Costill for dehydration (Dill and Costill, 1974 (link); Alis et al., 2015 (link)).

All the analytical measurements were performed at the time of enrolment using blind-coded samples (no name or personal identifiers). Peripheral venous blood samples were obtained from participants in the study in the fasting state at least twelve hours after the last meal. After allowing clotting for at least 30 min, the samples were centrifuged at 1600 ×g for 15 min. Besides, a whole-blood sample was obtained, and DNA was extracted.
Serum leptin, adiponectin, neurotensin, BDNF, and plasma IL-6, IL-8, TNF-α, LPS, and TLR-4 concentrations were measured in duplicate using commercially available sandwich enzyme-linked immunosorbent assay kits: Human Leptin ELISA—Diagnostic Biochem, Canada Inc., Ontario, Canada; Human Adiponectin ELISA, High Sensitivity—Biovendor GmbH, Heidelberg, Germany; ELISA kit for neurotensin—Cloud-Clone Corp., TX, USA; Human Free BDNF Elisa kit—R&D Systems Inc., MN, USA); Human IL-6 Quantikine ELISA, Human IL-8 Quantikine ELISA, and Human TNF-alpha Quantikine ELISA kits—BD Biosciences, Milan Italy; Lipopolysaccharide (LPS) ELISA kit—Cloud-Clone Corp., Katy, TX, USA; Human Toll-Like Receptor 4 (TLR-4) ELISA kit—Cloud-Clone Corp., Katy, TX, USA.