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.
Recombinant Human Brain Derived Neurotrophic Factor (BDNF).
Research Sub Category
Neurochemistry & Neurotrophins
Neuroinflammation & Pain
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.
~12-18kDa monomer, with a 28-30kDa homodimer in most samples.
Protein G Purified
Protein G Purified immunoglobulin. Lyophilized from PBS, no preservatives. Reconstitute with 500 μL of sterile distilled water.
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.
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
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