Analysis Note

ControlBrain tissue

Application

Research CategoryNeuroscience

Anti-Brain Derived Neurotrophic Factor Antibody detects level of Brain Derived Neurotrophic Factor & has been published & validated for use in ELISA, IH, FUNC & WB.

Immunohistochemistry: 1-5 µg/mL (see suggested protocol).

Immunoblotting: 1-5 µg/mL we recommend preparing the BDNF samples for western blot by the method of Semba-Katoh, R et.al (J. Neurochem. 69(1):34-42, 1997) because BDNF is not easily extracted in standard buffers. Dissected tissues should be homogenized in 10 volumes of 100mM phosphate buffer containing 1mM EDTA, 2M guanidine hydrochloride (pH7.2), and three protease inhibitors, 10mM N-ethylmaleimide, 0.36mM pepstatin, and 1mM PMSF. Homogenates are then sonicated and centrifuged at 46K x g for 30 minutes at 4°C. BDNF migrates as ~12-18kDa monomer, with a 28-30kDa homodimer in most samples.

ELISA: 1-5 µg/mL

Inhibition of biological activity in vitro: Complete inhibition of BDNF activity on embryonic chick sensory neurons occurs at 1-5 µg/mL. Biological inhibition protocols can be found in Finn et al. (J. Neurocytol. 15:169-176, 1986).
BIOLOGICAL ACTIVITY: Neutralizes BDNF, but not other neurotrophins.

Use neat for in vivo studies.

Optimal working dilutions must be determined by end user.

Research Sub CategoryNeurochemistry & NeurotrophinsNeuroinflammation & Pain

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Immunogen

Recombinant Human Brain Derived Neurotrophic Factor (BDNF).

Legal Information

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Physical form

Protein G Purified

Format: Purified

Protein G Purified immunoglobulin. Lyophilized from PBS, no preservatives. Reconstitute with 500 µL of sterile distilled water.

Specificity

Recognizes Brain Derived Neurotrophic Factor (BDNF). Neutralizes BDNF, but not other neurotrophins. By one site ELISA, less than 1% cross-reactivity against mouse NGF, recombinant human NT3 or neurotrophin 4. The antibody was tested on human and rat derived samples, but based on sequence similarity it is expected to react with mouse and other mammalian species. The peptide sequences of human, rat and mouse BDNF are identical.

Storage and Stability

Maintain lyophilized material at -20 to -70°C for up to 12 months after date of receipt. After reconstitution maintain at -20°C in undiluted aliquots for up to 6 months. Glycerol (1:1) can be added for additional stability. Avoid repeated freeze/thaw cycles.

Target description

~12-18kDa monomer, with a 28-30kDa homodimer in most samples.