Brain-derived neurotrophic factor, also known as BDNF, is a prosurvival factor induced by cortical neurons that is necessary for survival of striatal neurons in the brain. BDNF is a member of the "neurotrophin" family of growth factors, which are related to the NGF. It acts on certain neurons of the central nervous system and the peripheral nervous system, helping to support the survival of existing neurons, and encourage the growth and differentiation of new neurons and synapses. In the brain, it is active in the hippocampus, cortex, and basal forebrain—areas vital to learning, memory, and higher thinking. Post mortem analysis has shown lowered levels of BDNF in the brain tissues of people with Alzheimer's disease.
Principle of the Assay
This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-BDNF polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-BDNF polyclonal antibody was used as detection antibodies. The standards, test samples and biotin conjugated detection antibody were added to the wells subsequently, and wash with wash buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with wash buffer. TMB substrates were used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the BDNF amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of BDNF can be calculated.
Range: 31.2 pg/ml-2000 pg/ml
Sensitivity < 10 pg/ml
Storage and Expiration: Store at 2-8℃ for 6 months, or at -20℃ for 12 months.
Application: For quantitative detection of BDNF in Rabbit serum, plasma, body fluids, tissue lysates or cell culture supernatants.
1. One 96-well plate pre-coated with anti-Rabbit BDNF antibody
2. Lyophilized Rabbit BDNF standards: 2 tubes (10 ng/ tube)
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-Rabbit BDNF antibody (Concentrated): 130μl. Dilution: 1:100
5. Antibody diluent buffer: 12ml
6. Avidin-Biotin-Peroxidase Complex (ABC) (Concentrated): 130μl. Dilution: 1:100
7. ABC diluent buffer: 12ml
8. TMB substrate: 10ml
9. Stop solution: 10ml
10. Wash buffer (25X): 30ml
Note: Reconstitute standards and test samples with Kit Component 3.