Anti-BDNF Antibody

Brain-derived neurotrophic factor
    Cat #: ANT-010
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, m, r
    Immunogen
    Peptide (C)VLEKVPVSKQLK, corresponding to amino acid residues 166-178 of human BDNF (precursor) (Accession P23560).
    Accession (Uniprot) Number P23560
    Gene ID 627
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Mouse, rat and many other species - identical.
    Purity Affinity purified on immobilized antigen.
    Formulation Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.025% NaN3.
    Specificity The antibody is specific for BDNF; it does not crossreact with NGF, NT-3 or NT-4.
    Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
    Reconstitution 25 μl, 50 μl or 0.2 ml double distilled water (DDW), depending on the sample size.
    Antibody concentration after reconstitution 0.72 mg/ml.
    Storage after reconstitution The reconstituted solution can be stored at 4°C for up to 1 week. For longer periods, small aliquots should be stored at -20°C. Avoid multiple freezing and thawing. Centrifuge all antibody preparations before use (10000 x g 5 min).
    Control antigen storage before reconstitution Lyophilized powder can be stored intact at room temperature for 2 weeks. For longer periods, it should be stored at -20°C.
    Control antigen reconstitution 100 µl double distilled water (DDW).
    Control antigen storage after reconstitution -20°C.
    Preadsorption Control 1 μg peptide per 1 μg antibody.
    Standard quality control of each lot Western blot analysis.
    Applications: ic, ih, wb
    May also work in: ifc, ip
    Western blot
    Western blot analysis with Anti-BDNF Antibody:
    1,5. Recombinant human BDNF protein (#B-250).
    2,6. Recombinant mouse proBDNF protein (#B-240).
    3,7. Recombinant human beta-NGF protein (#N-245).
    4,8. Recombinant human Neurotrophin-3 (NT-3) protein (#N-260).
    Lanes 1-4: Anti-BDNF Antibody (#ANT-010), (1:200).
    Lanes 5-8: Anti-BDNF Antibody, preincubated with the control peptide antigen.
     
    Note that the antibody recognizes both BDNF and proBDNF but fails to recognize the closely related NGF and NT-3 neurotrophins.
    Immunohistochemistry
    Expression of BDNF in mouse cerebellum
    Immunohistochemical staining of mouse cerebellum with Anti-BDNF Antibody (#ANT-010). A. BDNF (red) appears in Purkinje cells (upward pointing arrows) and is distributed diffusely in the molecular layer (Mol) including in astrocytic fibers (downward pointing arrows). B. Staining of astrocytic fibers with glial fibrillary acidic protein (green) in the same section demonstrates the distribution of BDNF to neuronal as well as to astrocytic cellular components. C. Confocal merge of BDNF and GFAP.
    Immuno-colocalization of Kir6.2 and BDNF in mouse hippocampus.
    Immunohistochemical staining of immersion-fixed, free floating mouse brain frozen sections using Guinea pig Anti-Kir6.2 Antibody (#AGP-067), (1:300) and Anti-BDNF Antibody (#ANT-010), (1:300). A. Kir6.2 staining (red) appears in the dentate gyrus granule layer (G) and in hilar interneurons (arrows). B. BDNF staining (green) in the same section appears in the dentate gyrus granule layer (G) and in hilar interneurons (arrows). C. Merge of the two images reveals colocalization of Kir6.2 and BDNF in hippocampal interneurons.
    Immunocytochemistry
    BDNF transfected striatal cells (1:50) (del Toro, D. et al.(2006) J. Neurosci. 26, 12748.).
    References
    1. Chao, M.V. (2003) Nature Rev. Neurosci. 4, 299.
    2. Blum, R. and Konnerth, A. (2005) Physiology 20, 70.
    3. Kalb, R. (2005) Trends Neurosci. 28, 5.
    4. Lu, B. et al. (2005) Nature Rev. Neurosci. 6, 603.
    Scientific background

    Brain-derived neurotrophic factor (BDNF) is a member of the neurotrophin family of growth factors which includes nerve growth factor (NGF), neurotrophin-3 (NT-3) and neurotrophin-4/5 (NT-4/5).

    All neurotrophins are synthesized as preproneurotrophin precursors that are subsequently processed within the intracellular transport pathway to yield proneurotrophins that are further processed to generate the mature form. The mature form of BDNF is a non-covalent stable homodimer that can be secreted in both constitutive and regulated pathways.

    BDNF conveys its activity by binding to two classes of receptors, a member of the Trk receptor tyrosine kinase family (TrkB) and the pan-neurotrophin receptor p75NTR. Binding of BDNF to the TrkB receptor triggers ligand-induced dimerization and autophosphorylation of tyrosine residues. This activates various signaling cascades such the MAPK, PI3K and PLCγ pathways that are involved in cell growth, survival and differentiation of neurons in the central and peripheral nervous system.  Interestingly, recent evidence suggests that BDNF may influence target cell function via ion channel modulation. Ion channel activity in the target cells can be modulated by a TrkB-mediated mechanism that has not yet been determined. BDNF is able to block both KV1.3 and AMPA-subtype glutamate ion channel currents in sensory neurons, while it can induce activation of the TRPC3 cation channel in neurons and of the NaV1.9 Na+ channel in hippocampal neurons. These newly recognized BDNF actions underlie its “rapid” neuronal functions that include changes in neuronal  excitability, plasticity and synaptic transmission.

    Application key:

    CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, IFC- Indirect flow cytometry, IHC- Immunohistochemistry, IP- Immunoprecipitation, LCI- Live cell imaging, N- Neutralization, WB- Western blot

    Species reactivity key:

    H- Human, M- Mouse, R- Rat
    Image & Title:

    Anti-BDNF Antibody
    Expresssion of BDNF in mouse brain
    Immunohistochemical staining of mouse brain sections using Anti-BDNF Antibody (#ANT-010). BDNF staining (purple) is expressed in astrocytes and co-localizes with GFAP (blue) but not with neurofilament-1 (NF-L), (green).Adapted from Fulmer, C.G. et al. (2014) J. Neurosci. 34, 8186. with permission of the Society for Neuroscience.

    Last update: 05/11/2018

    Alomone Labs is pleased to offer a highly specific antibody directed against an epitope of human brain-derived neurotrophic factor. Anti-BDNF Antibody (#ANT-010) can be used in western blot, immunocytochemistry, and immunohistochemistry applications. It has been designed to recognize BDNF from rat, human, and mouse samples. The antibody is specific for BDNF; it does not crossreact with NGF, NT-3 or NT-4.

    For research purposes only, not for human use
    Citations
    Western blot citations
    1. Rat brain lysate (1:1000).
      Hovens, I.B. et al. (2015) Am. J. Physiol. 309, R148.
    2. Mouse brain lysate (1:200).
      Duarte-Neves, J. et al. (2015) Hum. Mol. Genet. 24, 5451.
    3. Human SH-SY5Y and rat primary cortical neuron lysates (1:1000).
      Shin, M.K. et al. (2014) Neuropharmacology 77, 414.
    4. Mouse brain lysate (1:2000).
      Shin, M.K. et al. (2014) Neurobiol. Aging. 35, 990.
    5. Rat lamina terminalis lysate (1:200).
      Clayton, S.C. et al. (2014) Am. J. Physiol. 306, R908.
    6. Rat brain lysates (1:500).
      Tian, X. et al. (2013) PLoS ONE 8, e69252.
    Immunohistochemistry citations
    1. Mouse retina sections (1:200).
      Benhar, I. et al. (2016) EMBO J. 35, 1219.
    2. Mouse brain sections (1:200).
      Wattananit, S. et al. (2016) J. Neurosci. 36, 4182.
    3. Rat brain sections (8 µg/ml).
      Louveau, A. et al. (2015) Glia 63, 2298.
    4. Mouse brain sections.
      Fulmer, C.G. et al. (2014) J. Neurosci. 34, 8186.
    Immunocytochemistry citations
    1. BDNF transfected striatal cells (1:50).
      Del Toro, D. et al. (2006) J. Neurosci. 26, 12748.
    More product citations
    1. Demel, C. et al. (2011) J. Neuroinflammation. 8, 7.
    2. Poblete-Naredo, I. et al. (2011) Neurochem. Int. 59, 1133.
    3. España, J. et al. (2010) J. Neurosci. 30, 9402.
    4. Kong, Li. et al. (2010) Neurobiol. Dis. 38, 446.
    5. Taliaz, D. et al. (2010) Mol Psychiatry 15, 80.
    6. Lewitus, G.M. et al. (2008) Brain Behav. Immun. 22, 1108.
    7. Fiszman, M.L. et al. (2005) J. Neurosci. 25, 2024.
    Related Products
      • Antibodies for immuno-colocalization experiments
        1. Guinea pig Anti-BDNF Antibody (#AGP-021) is raised in guinea pig and can be used in immuno-colocalization studies in conjunction with any of our antibodies raised in rabbit. This antibody has been raised against the same epitope as #ANT-010.