Western blot analysis of extracts from 293T cells, untransfected or transfected with human BDNF protein (upper, lanes 1 and 2), or treated with human recombinant Human Brain-Derived Neurotrophic Factor (BDNF) #3897 protein, at the indicated levels, using BDNF Antibody (upper, lanes 3 and 4). Myc-Tag Antibody #2272 was used to show plasmid expression (middle). α-Actinin (D6F6) XP® Rabbit mAb #6487 was used as a loading control (lower).
Western blot analysis was performed with different amounts of the mouse recombinant BDNF protein using BDNF Antibody.
|MW (kDa)||Precursor 28 - Mature 12-14|
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 10
BDNF Antibody recognizes human and mouse recombinant BDNF, and exogenously expressed BDNF.
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding amino acid His129 of human BDNF protein.
Neurotrophins are comprised of at least four family members including NGF, BDNF, NT-3 and NT-4 and all are known to influence growth, development, differentiation and survival of neurons (1). Proneurotrophins bind to p75NTR but not to the family of Trk receptor tyrosine kinases (Trk) and following maturation, BDNF binds and activates TrkB. Trk receptors in turn activate three major signaling pathways: (a) Ras-MAPK signaling, which promotes neuronal differentiation and neurite outgrowth, (b) PI3 Kinase-Akt signaling, which promotes survival and growth of neurons, and (c) PLC-γ1-PKC signaling, which promotes synaptic plasticity (2). BDNF is a major regulator of transmission and plasticity at adult synapses. Moreover, the precursor proBDNF and the mature protein mBDNF drive opposite effects on long-term potentiation and long-term depression (3). BDNF has also been implicated in body weight regulation and activity: heterozygous BDNF knockout mice are hyperphagic, obese, and hyperactive (4).
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