Anti-proBDNF-ATTO-488 Antibody

Brain-derived neurotrophic factor precursor
A Rabbit Polyclonal Antibody to proBDNF Conjugated to the Fluorescent Dye ATTO-488
    Cat #: ANT-006-AG
    Alternative Name Brain-derived neurotrophic factor precursor
  • Lyophilized Powder
  • Antigen Incl.
  • Type: Polyclonal
    Host: Rabbit
    Reactivity: h, m, r
      • Peptide (C)DEDQKVRPNEENNKDAD, corresponding to amino acid residues 72-88 of human BDNF (precursor) (Accession P23560). Prodomain of the BDNF protein.
        Anti-proBDNF-ATTO-488 Antibody
    Accession (Uniprot) Number P23560
    Gene ID 627
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Mouse, rat - 16 /17 amino acid residues identical.
    RRID AB_10557039.
    Purity Affinity purified on immobilized antigen.
    Form Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN3.
    Isotype Rabbit IgG.
    Label ATTO-488. Maximum absorption 501 nm; maximum fluorescence 523 nm. The fluorescence is excited most efficiently in the 480 – 515 nm range. This label is analogous to the well known dye fluorescein isothiocyanate (FITC) and can be used with filters typically used to detect FITC.
    Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
    Reconstitution 50 µl double distilled water (DDW).
    Antibody concentration after reconstitution 1 mg/ml.
    Storage after reconstitution The reconstituted solution can be stored at 4°C, protected from the light, 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 × g 5 min).
    Negative 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.
    Negative control antigen reconstitution 100 µl double distilled water (DDW).
    Negative 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 (unlabeled antibody, #ANT-006), and immunohistochemistry (labeled antibody).
    Applications: fc, if, ih
    May also work in: ic*
      • Anti-proBDNF-ATTO-488 Antibody
        Expression of proBDNF in rat DRG
        Immunohistochemical staining of rat dorsal root ganglia (DRG) frozen sections using Anti-proBDNF-ATTO-488 Antibody (#ANT-006-AG), (1:50), (green). Staining is present in DRG neurons. Hoechst 33342 is used as the counterstain (blue).
      • Human isolated platelets (Azoulay, D. et al. (2014) Br. J. Haematol. 164, 451.).
    References
      • Brain derived neurotrophic factor (BDNF) is a member of the neurotrophin family of growth factors that 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.

        Until recently, the functional role of the neurotrophin prodomains were thought to include assistance in the correct folding of the mature protein and the sorting of the neurotrophins into the constitutive or regulated secretory pathway. However, a growing body of evidence suggests that the uncleaved proneurotrophin precursors can be secreted from cells and that they may mediate different biological functions.

        The functional importance of the prodomain of BDNF was recently demonstrated in a study showing that a polymorphism that replaces valine for methionine at position 66 of the prodomain, is associated with memory defects and abnormal hippocampal function in humans. Another recent study showed that the regulated extracellular cleavage of proBDNF to mature BDNF by plasmin is necessary for establishing late-phase long-term potentiation (L-LTP), a process that involves long-lasting changes in the structure and function of hippocampal synapses. Finally, proBDNF was shown to be decreased in the brains of patients suffering from Alzheimer’s disease.

        Mature BDNF binds to the specific tyrosine kinase receptor TrkB and to p75NTR, a member of the TNF receptor superfamily. proBDNF can similarly bind both receptors although it appears to have a greater affinity for the p75NTR receptor.

    Application key:

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

    Species reactivity key:

    H- Human, M- Mouse, R- Rat
    Last update: 07/11/2019

    Anti-proBDNF Antibody (#ANT-006) is a highly specific antibody directed against the prodomain region of human proBDNF. The antibody can be used in western blot, immunoprecipitation, and immunohistochemistry applications. It has been designed to recognize proBDNF from human, mouse, and rat samples. The antibody does not cross react with mature BDNF, pro- and mature NGF or mature NT-3.

    Anti-proBDNF-ATTO-488 Antibody (#ANT-006-AG) is directly labeled with an ATTO-488 fluorescent dye. ATTO dyes are characterized by strong absorption (high extinction coefficient), high fluorescence quantum yield, and high photo-stability. The ATTO-488 label is analogous to the well known dye fluorescein isothiocyanate (FITC) and can be used with filters typically used to detect FITC. Anti-proBDNF-ATTO-488 Antibody is especially suited for experiments requiring simultaneous labeling of different markers.

    For research purposes only, not for human use
    Citations
      • Human isolated platelets.
        Azoulay, D. et al. (2014) Br. J. Haematol. 164, 451.
    Related Products
        • Guinea pig Anti-proBDNF Antibody (#AGP-032) 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-006.