Guinea pig Anti-proBDNF Antibody

Brain-derived neurotrophic factor precursor
Cat #: AGP-032
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Guinea pig
    Reactivity: h, m
    May also work in: r
    Immunogen
    Peptide (C)DEDQKVRPNEENNKDAD, corresponding to amino acid residues 72-88 of human BDNF (precursor) (Accession P23560). Pro-domain of the BDNF protein.
    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.
    Purity Affinity purified on immobilized antigen.
    Formulation Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN3.
    Specificity The antibody is specific for proBDNF, it does not crossreact with proNGF, proNT-3 or mature BDNF.
    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 0.8 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: ih, wb
    May also work in: cbe, ic, ifc, ip
    Western blot
    Western blot analysis of 50 ng of each Recombinant human Val66Met proBDNF (cleavage resistant) protein (#B-456) (lanes 1 and 4), Recombinant mouse proBDNF protein (#B-240) (lanes 2 and 5) and Recombinant human proBDNF protein (#B-257) (lanes 3 and 6):
    1-3. Guinea pig Anti-proBDNF Antibody (#AGP-032), (1:200).
    4-6. Guinea pig Anti-proBDNF Antibody, preincubated with the control peptide antigen.
    Following a broad screen of secondary antibodies, the following was used for this application: #106-035-006 (Jackson ImmunoResearch).
    Immunohistochemistry
    Expression of proBDNF in rat spinal cord
    Immunohistochemical staining of immersion-fixed, free floating rat spinal cord frozen sections using Guinea pig Anti-proBDNF Antibody (#AGP-032), (1:1000). proBDNF (red) is detected in the superficial layer (arrow) of the dorsal horn of the spinal cord. The myelinated columns are devoid of staining. Cell nuclei are stained with DAPI (blue).
    References
    1. Lee R. et al. (2001) Science 294, 1945.
    2. Egan, M.F. et al. (2003) Cell 112, 257.
    3. Pang, P.T. et al. (2004) Science 306, 487.
    4. Michalski, B. et al. (2003) Mol. Brain Res. 111, 148.
    Scientific background

    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, IHC- Immunohistochemistry, IP- Immunoprecipitation, LCI- Live cell imaging, N- Neutralization, WB- Western blot

    Species reactivity key:

    H- Human, M- Mouse, R- Rat
    Last update: 05/02/2019

    Alomone Labs is pleased to offer a highly specific antibody directed against the prodomain region of human proBDNF. Guinea pig Anti-proBDNF Antibody (#AGP-032), raised in guinea pig, can be used in western and immunohistochemistry applications. In addition, the antibody is an excellent tool for easy immuno-colocalization studies. It has been designed to recognize proBDNF from mouse, rat and human samples. The antigen used to immunize guinea pigs is the same as Anti-proBDNF Antibody (#ANT-006) raised in rabbit. Our line of guinea pig antibodies enables more flexibility with our products such as immuno-colocalization studies, immunoprecipitation, etc.

    For research purposes only, not for human use
    Citations
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