Anti-TrkB (extracellular) Antibody

Neurotrophic tyrosine kinase receptor type 2, NTRK2, BDNF/NT-3 growth factors receptor
    Cat #: ANT-019
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, m, r
    Immunogen
    Peptide (C)RLEPNSIDPENITE, corresponding to amino acid residues 57-70 of rat TrkB (Accession Q63604). Extracellular domain.
    Accession (Uniprot) Number Q63604
    Gene ID 25054
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Mouse, human - 13/14 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, 0.025% NaN3.
    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: ic, ifc, ih, lci, wb
    May also work in: ip
    Western blot
    Western blot analysis of rat brain membranes (lanes 1 and 3) and HEK-TrkB transfected cell lysates (lanes 2 and 4):
    1,2. Anti-TrkB (extracellular) Antibody (#ANT-019), (1:200).
    3,4. Anti-TrkB (extracellular) Antibody, preincubated with the control peptide antigen.
    Western blot analysis of mouse brain lysates:
    1. Anti-TrkB (extracellular) Antibody (#ANT-019), (1:200).
    2. Anti-TrkB (extracellular) Antibody, preincubated with the control peptide antigen.
    Immunohistochemistry
    Expression of TrkB in rat DRG
    Immunohistochemical staining of rat dorsal root ganglia (DRG) frozen sections using Anti-TrkB (extracellular) Antibody (#ANT-019), (1:100). TrkB (red) is expressed in DRG neurons. Hoechst 33342 is used as the counterstain.
    Indirect flow cytometry
    Indirect flow cytometry analysis of live intact HL-60 (human promyelocytic leukemia cells) cell line:
    ___ Cells + goat-anti-rabbit-FITC.
    ___ Cells + Anti-TrkB (extracellular) Antibody (#ANT-019), (1:25) + goat-anti-rabbit-FITC.
    The control antigen is not suitable for this application.
    Live cell imaging / Immunocytochemistry
    Expression of TrkB in rat PC12 cells
    Immunocytochemical staining of live intact rat pheochromocytoma PC12 cells. A. Extracellular staining of cells with Anti-TrkB (extracellular) Antibody (#ANT-019) (1:50), followed by goat anti-rabbit-AlexaFluor-594 secondary antibody (red). B. Live view of the cells. C. Merge of the two images.
    References
    1. Webster, N.J.G. and Pirrung, M.C. (2008) BMC Neurosci. 9, S1.
    2. Quartu, M. et al. (2003) Int. J. Dev. Neurosci. 21, 309.
    3. Aoki, C. et al. (2000) J. Neurosci. Res. 59, 454.
    4. Yan, Q. et al. (1997) J. Comp. Neurol. 378, 135.
    5. Patapoutian, A. and Reichardt, L.F. (2001) Curr. Opin. Neurobiol. 11, 272.
    6. Schecterson, L.C. and Bothwell, M. (2010) Develop. Neurobiol. 70, 332.
    7. Grimes, M.L. et al. (1996) J. Neurosci. 16, 7950.
    Scientific background

    BDNF and NT-4 belong to the neurotrophin family which also includes NGF and NT-3. These neurotrophins bind two groups of receptors. The p75NTR receptor is common to all four neurotrophins and is a member of the tumor necrosis factor receptor family. The tropomyosin-related kinase (Trk) receptors are receptor tyrosine kinases (RTKs) and three receptors form this family: TrkA, TrkB, and TrkC1.

    As mentioned above, the p75NTR receptor binds to all neurotrophins with similar affinities while the Trk receptors are the ones to display the selectivity for the neurotrophins. TrkA is activated by NGF binding, TrkB by that of BDNF and NT-4, while TrkC is stimulated by the binding of NT-31.

    All three Trk receptors are highly expressed in the mammalian brain in very distinct regions and are also expressed in the peripheral nervous system2-4. Cholinergic neurons in the basal forebrain exclusively express TrkA, while TrkB and TrkC are highly expressed in the hippocampus. Motor and sensory neurons in the peripheral nervous system express Trk receptors. Interestingly, Trk receptors are not essential for development, but knockout mice die shortly after birth. Indeed, TrkB-deficient mice demonstrate a significant decrease in motor neurons and synaptogenesis1.

    Trk receptors have many motifs in the extracellular region, including cell-adhesion domains, three tandem leucine rich motifs flanked by two clusters of cysteines. In the membrane proximal region of the receptor there are also two immunoglobulin-like domains5. The binding of neurotrophins to Trk receptors promotes receptor dimerization resulting in kinase activation. Activated Trk receptors then phosphorylate a cascade of signaling molecules including the Ras/ERK, PI3K/Akt pathways and PLC-g1. Activated Trk receptors also create internal docking sites for other signaling adaptor proteins to bind to5. Splice variants of TrkA, TrkB and TrkC have been observed. These splice isoforms are mainly affected in the tyrosine kinase domain of the receptor lying in the cytoplasm. Endocytosis is an important signaling trait of Trk receptors.

    Following neurotrophin binding to the Trk receptor, the receptor complex is then internalized via endocytosis in order to terminate signaling. However, in the axonal compartment of neurons the internalization process of the neurotrophin complexed to the receptor is part of the signaling process and is important for activating transcription processes in the nucleus6,7.

    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: 29/11/2018

    Anti-TrkB (extracellular) Antibody (#ANT-019) is a highly specific antibody directed against an epitope of the rat protein. The antibody can be used in western blot, immunohistochemistry, indirect flow cytometry, and live cell imaging applications. It has been designed to recognize TrkB from rat, mouse, and human samples.

    For research purposes only, not for human use
    Citations
    Western blot citations
    1. Rat brain lysate (1:400).
      Schaich, C.L. et al. (2016) Auton. Neurosci. 197, 1.
    Immunohistochemistry citations
    1. Rat adrenal glands (1:100).
      Scott, A.L. et al. (2015) J. Physiol. 593, 3281.
    Immunocytochemistry citations
    1. Rat MAH chromaffin-derived cells.
      Scott, A.L. et al. (2015) J. Physiol. 593, 3281.
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