Quantikine human bdnf kit

For the BDNF expression studies, hippocampi were collected from mice sacrificed by cervical dislocation followed by anesthesia with CO₂. Samples were frozen on dry ice, and stored at –70°C until use. The BDNF expression was determined using BDNF ELISA (Quantikine human BDNF kit, R&D Systems) as described previously (Louhivuori et al., 2011 (link)).

Plasma BDNF level was measured in blood samples taken within a 2-h interval in the morning, between 9 AM and 11 AM, after patients had fasted for at least 8 h. Ten milliliters of whole blood were withdrawn from the antecubital vein of each patient into a vacuum tube containing ethylenediamine tetraacetic acid (EDTA) (Greiner Bio-One Vacuette; Santa Cruz Biotechnology, Santa Cruz, CA, USA), and the blood sample was kept on ice for up to 30 min. To isolate plasma, whole blood was then centrifuged at 3000 × g for 15 min at 4 °C and immediately stored at −80 °C. A BDNF kit (Quantikine Human BDNF kit; R&D Systems, Minneapolis, MN, USA) and an enzyme-linked immunosorbent assay (ELISA) reader (Spectra-Max-M2; Molecular Devices, Sunnyvale, CA, USA) were used to analyze the level of plasma BDNF, the minimum detectable dose of which is typically < 80 pg/ml for humans. All samples were assayed in duplicate by laboratory staff blind to patient status.

At baseline, 10 milliliters of venous blood were collected from each participants. DNA was extracted from the lymphocytes of the blood sample. The BDNF Val66Met polymorphism was genotyped using a modified protocol22 (link). All samples were double-checked to keep genotype error less than 5%.
Blood samples were collected between 9 am and 11 am after 8 hours of fasting at baseline and endpoint to measure Plasma BDNF level. Blood sample was drawn into a vacuum tube containing ethylenediamine tetraacetic acid (EDTA) (Greiner Bio-One Vacuette; Santa Cruz Biotechnology, Santa Cruz, CA) then kept on ice for up to 30 minutes. At 4 °C, the whole blood was centrifuged at 3000 g for 15 minutes to isolate plasma; then stored at −80 °C. A BDNF kit (Quantikine Human BDNF kit; R&D Systems, Minneapolis, MN), and an enzyme-linked immunosorbent assay (ELISA) reader (SpectraMax-M2; Molecular Devices, Sunnyvale, CA) with minimum detectable dose of 80 pg/ml were used to assess the level of plasma BDNF. All samples were analyzed twice.

Plasma BDNF levels were measured using a BDNF Kit (Quantikine Human BDNF kit; R&D Systems, Minneapolis, MN, USA) (www.RnDSystems.com) and an ELISA reader with a minimum detectable dose of 80 pg/mL (SpectraMax-M2; Molecular Devices, Sunnyvale, CA, USA). All samples were analyzed twice. The cytokine ELISA kits used were also purchased from R&D systems and included a Human CRP Quantikine ELISA Kit, a Human TNF-alpha Quantikine ELISA Kit, and a Human IL-8 Quantikine ELISA Kit for the assessment of plasma levels of CRP, TNF-α, and IL-8, respectively.