• Strip plates and additional reagents allow for use in multiple experiments
• Quantitative protein detection
• Establishes normal range
• The best products for confirmation of antibody array data

The BDNF ELISA employs an antibody specific for human BDNF coated on a 96-well plate. Standards and samples are pipetted into the wells and BDNF present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human BDNF antibody is added. After washing away unbound biotinylated antibody, HRPconjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of BDNF bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

Brain-derived neurotrophic factor (BDNF) is found in neurons of the central nervous system and is a member of the nerve growth factor family of proteins. It is expressed predominantly in hippocampus, cortex, and synapses of the basal forebrain. BDNF selectively supports the survival of primary sensory neurons and retinal ganglia. It supports survival and differentiation of certain cholinergic neurons and also some dopaminergic neurons in vitro. The factor inhibits the normal cell death of embryonic chick motor neurons. Expression of the BDNF gene is reduced in Alzheimer's, Parkinson's, and Huntington's disease patients. It may play a role in the regulation of the stress response and in the biology of mood disorders.

• Pre-Coated 96-well Strip Microplate
• Wash Buffer
• Stop Solution
• Assay Diluent(s)
• Lyophilized Standard
• Biotinylated Detection Antibody
• Streptavidin-Conjugated HRP
• TMB One-Step Substrate

• Distilled or deionized water
• Precision pipettes to deliver 2 µL to 1 µL volumes
• Adjustable 1-25 µL pipettes for reagent preparation
• 100 µL and 1 liter graduated cylinders
• Tubes to prepare standard and sample dilutions
• Absorbent paper
• Microplate reader capable of measuring absorbance at 450nm
• Log-log graph paper or computer and software for ELISA data analysis

1. Prepare all reagents, samples and standards as instructed in the manual.
2. Add 100 µL of standard or sample to each well.
3. Incubate 2.5 h at RT or O/N at 4°C.
4. Add 100 µL of prepared biotin antibody to each well.
5. Incubate 1 h at RT.
6. Add 100 µL of prepared Streptavidin solution to each well.
7. Incubate 45 min at RT.
8. Add 100 µL of TMB One-Step Substrate Reagent to each well.
9. Incubate 30 min at RT.
10. Add 50 µL of Stop Solution to each well.
11. Read at 450 nm immediately.