Brain-Derived Neurotrophic Factor (BDNF) ELISA Assay Kit

$550.00

The Eagle Biosciences BDNF (Brain-Derived Neurotrophic Factor) ELISA Assay Kit is an Enzyme Immunoassay kit for the quantification of Human BDNF in serum, plasma or cell culture supernatants. The Brain-Derived Neurotrophic Factor (BDNF) ELISA Assay Kit is for research use only and should not be used for diagnostic procedures.

SKU: ARG81251 Categories: , ,

Brain-Derived Neurotrophic Factor (BDNF) ELISA Assay Kit

For Research Use Only

Size: 1×96 wells
Sensitivity: 7 pg/ml
Dynamic Range: 15.625-1000 pg/ml
Incubation Time: 3.5 hours
Sample Type: Serum, Plasma, culture supernatants
Sample Size: 100 µL

Additional Information

Assay Background


The protein encoded by this gene is a member of the nerve growth factor family. It is induced by cortical neurons, and is necessary for survival of striatal neurons in the brain. Expression of this gene is reduced in both Alzheimer’s and Huntington disease patients. This gene may play a role in the regulation of stress response and in the biology of mood disorders. Multiple transcript variants encoding distinct isoforms have been described for this gene. [provided by RefSeq, Jan 2009]

During development, BDNF promotes the survival and differentiation of selected neuronal populations of the peripheral and central nervous systems. Participates in axonal growth, pathfinding and in the modulation of dendritic growth and morphology. Major regulator of synaptic transmission and plasticity at adult synapses in many regions of the CNS. The versatility of BDNF is emphasized by its contribution to a range of adaptive neuronal responses including long-term potentiation (LTP), longterm depression (LTD), certain forms of short-term synaptic plasticity, as well as homeostatic regulation of intrinsic neuronal excitability.

Assay Principle


The Eagle Biosciences BDNF (Brain-Derived Neurotrophic Factor) ELISA Assay Kit employs the quantitative sandwich enzyme immunoassay technique. An antibody specific for BDNF has been pre-coated onto a microtiter plate. Standards or samples are pipetted into the wells and any BDNF present is bound by the immobilized antibody. After washing away any unbound substances, a biotin-conjugated antibody specific for BDNF is added to each well and incubate. Following a washing to remove unbound substances, streptavidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After washing away any unbound antibody-enzyme reagent, a substrate solution (TMB) is added to the wells and color develops in proportion to the amount of BDNF bound in the initial step. The color development is stopped by the addition of acid and the intensity of the color is measured at a wavelength of 450nm ±2nm.The concentration of BDNF in the sample is then determined by comparing the O.D of samples to the standard curve.

Assay Procedure


  1. Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal it.
  2. Add 100 μl of standards, samples and zero controls (Standard/Sample diluent buffer) into wells. Incubate for 1.5 h at 37 °C.
  3. Aspirate each well and wash, repeating the process four times for a total five washes. Wash by filling each well with 1× Wash Buffer (350 μl) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating, decanting or blotting against clean paper towels.
  4. Add 100 μl 1X Antibody conjugate into each well. Cover wells and incubate for 1 hour at 37 °C.
  5. Aspirate each well and wash as step 3.
  6. Add 100 μl of 1X HRP-Streptavidin solution to each well. Cover wells and incubate for 30 minutes at 37 °C.
  7. Aspirate each well and wash as step 3.
  8. Add 100 μl of TMB Reagent to each well. Incubate for 10-20 minutes at 37°C in dark.
  9. Add 100 μl of Stop Solution to each well. The color of the solution should change from blue to yellow. Gently tap the plate to ensure thorough mixing
  10. Read the OD with a microplate reader at 450nm immediately. (optional: read at 610-650 nm as the reference wave length).

Typical Standard Curve


BDNF ELISA Standard Curve

Manual

Product Manual