Fasting blood was collected between 07:00 and 07:30 a.m. , the morning after PSG monitoring using a serum separator tube. Serum samples were prepared by centrifugation at 3200 × g for 10 minutes at 4°C and stored at −80°C until assaying. Serum BDNF and NfL levels were measured using an enzyme‐linked immunosorbent assay kit according to the manufacturer's instructions (Ray Bio Human BDNF ELISA Kit Cat: ELH‐BDNF Lot: 0515200106, Abbexa abx258398 Human NfL ELISA Kit Lot: E2002668G). The absorbance was measured at 450 nm using a microplate reader to determine BDNF (ng/mL) and NfL (pg/mL) concentrations.
Human bdnf elisa kit
Manufactured by Abbexa
The Human BDNF ELISA Kit is a quantitative sandwich enzyme immunoassay designed for the detection of human Brain-Derived Neurotrophic Factor (BDNF) in serum, plasma, cell culture supernatants, and other biological fluids.
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2 protocols using human bdnf elisa kit
1
Measuring Serum BDNF and NfL Levels
2
Evaluating Human BDNF Expression in P301L Mouse Brains
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ELISA, western blot and immunohistochemistry assays were performed to examine the expression of human BDNF in the brains of P301L mice after intraventricular injection of AAV-BDNF. Fresh left hemisphere was weighed and homogenized in lysate buffer. The homogenates were centrifuged at 10 000 g for 10 min at 4 °C, and the resultant supernatant was collected and subjected to ELISA assays according to the manufacturer's protocols (human BDNF ELISA kit, Abbexa). In addition, the levels of total BDNF including endogenous mouse BDNF and human BDNF in brain homogenates were also detected by western blot analysis using anti-BDNF antibody (Abcam, ab72439, detecting both mouse and human BDNF). Coronal sections from the right hemisphere were immunostained for microglia (anti-CD45 antibody, Millipore, Bedford, MA, USA), astrocyte (anti-glial fibrillary acidic protein antibody, Millipore), neurons (anti-NeuN antibody, Abcam), GFP (anti-GFP antibody, Abcam) and human BDNF (anti-human BDNF antibody, R&D systems, Minneapolis, MN, USA) to identify localization and levels of human BDNF expression in P301L mouse brains after AAV-BDNF injection.
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