Anti-p75 NGF Receptor (extracellular) Antibody

p75NTR, p75 Neurotrophin receptor, Low affinity NGF receptor, TNFR superfamily member 16, TNFRSF16, CD271
Cat #: ANT-007
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, m, r
    Peptide CEEIPGRWITRSTPPE, corresponding to amino acid residues 188-203 of human p75NTR (Accession P08138). Extracellular, stalk domain.
    Accession (Uniprot) Number P08138
    Gene ID 4804
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Mouse - identical; rat - 15/16 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, ip, lci, wb
    Western blot
    Western blot analysis of rat brain membranes:
    1. Anti-p75 NGF Receptor (extracellular) Antibody (#ANT-007), (1:200).
    2. Anti-p75 NGF Receptor (extracellular) Antibody, preincubated with the control peptide antigen.
    Western blot analysis of human melanoma cells A875:
    1. Anti-p75 NGF Receptor (extracellular) Antibody (#ANT-007), (1:200).
    2. Anti-p75 NGF Receptor (extracellular) Antibody, preincubated with the control peptide.
    Western blot analysis of normal rat tissue (right) and in human cancer cell lines (left):
    p75NTR is visualized with Anti-p75 NGF Receptor (extracellular) Antibody (#ANT-007), (1:200). Note that human cancer cell lines from hemapoietic origin show high p75NTR expression, while cell lines from prostate and colon cancer origin show lower levels. Interestingly, p75NTR from rat (right blot and the C6 cell line) and human (left blot) samples run with a different apparent MW, probably due to species-specific differential glycosylation.
    Immunoprecipitation of 3T3/p75NTR transfected cells:
    1. Cell lysate + protein A beads.
    2. Cell lysate + protein A beads + pre-immune rabbit serum.
    3. Cell lysate + protein A beads + Anti-p75 NGF Receptor (extracellular) Antibody (#ANT-007).
    4. Cell lysate.

    Red arrows indicate NGFR p75 while the black arrow shows the IgG heavy chain.
    Immunoblot was performed with the Anti-p75 NGF Receptor (extracellular) Antibody.
    Expression of p75NTR in rat brain
    Immunohistochemical staining of rat brain with Anti-p75 NGF Receptor (extracellular) Antibody (#ANT-007). A. Cells in the diagonal band are stained positive for p75NTR. B. Staining of the same section with goat anti-ChAT confirms that p75NTR staining is specific to cholinergic neurons.
    Expression of p75NTR in rat brain
    Immunohistochemical staining of rat brain with Anti-p75 NGF Receptor (extracellular) Antibody (#ANT-007). A. Cells in the nucleus basalis mangocellularis are stained positive for p75NTR. B. Staining of the same section with goat anti-ChAT confirms that the p75NTR staining is specific to cholinergic neurons.
    Indirect flow cytometry
    Indirect flow cytometry analysis of A875 human melanoma cells
    The control antigen is not suitable for this application.
    Live cell imaging / Immunocytochemistry
    A875 human live melanoma cells lines (1:100-1:200).
    1. Hempstead, B.L. (2002) Curr. Opin. Neurobiol. 12, 260.
    2. Chao, M.V. (2003) Nat. Rev. Neurosci. 4, 299.
    3. Barker, P.A. (2004) Neuron 42, 529.
    Scientific background

    The p75 neurotrophin receptor (p75NTR) is a member of the TNF receptor superfamily. p75NTR, like all the superfamily members is a type I transmembrane protein with tandem cysteine-rich domains in the extracellular portion and a intracellular death domain. p75NTR binds its ligands as a homodimer but can also form heterodimers with other receptors such as TrkATrkBTrkCNogo receptor and sortilin. The precise multimeric receptor complex formed between p75NTR and the other receptors will determine the ligand being recognized (see below) and the biological response to its binding.

    As it name implies, p75NTR binds all the neurotrophins (NGFBDNFNT-3 and NT-4) with similar nM affinities. Co-expression of p75NTR with the Trk receptors enhances the ability to bind and respond to the specific neurotrophin and induces cell survival. On the other hand, it has recently been demonstrated that p75NTR binds with high affinity to the unprocessed form of NGF (proNGF) probably as a complex with sortilin and this leads to cell death by apoptosis. Finally, a multimeric complex of p75NTR and Nogo receptor binds myelin proteins such as Nogo-66, MAG and OmgP resulting in inhibition of axonal growth.

    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: 30/12/2018

    Anti-p75 NGF Receptor (extracellular) Antibody (#ANT-007) is a highly specific antibody directed against an extracellular epitope of the human protein. The antibody can be used in western blot, immunoprecipitation, immunohistochemistry, live cell imaging and indirect flow cytometry applications. It has been designed to recognize p75NTR from human, mouse and rat samples.

    For research purposes only, not for human use
    Live cell imaging citations
    1. Rat PC12 cells (also used to observe internalization of the receptor).
      Escudero, C.A. et al. (2014) J. Cell Sci. 127, 1966.
    More product citations
    1. Civenni, G. et al. (2011) Cancer Res. 71, 3098.
    2. D'Onofrio, M. et al. (2011) PLoS One. 6, e20839.
    3. Wang, Y-J. et al. (2011) J. Neurosci. 31, 2292.
    4. Chakravarthy, B. et al. (2010) Biochem. Biophys. Res. Com. 401, 458.
    5. Curchoe, C.L. et al. (2010) PLoS One. 5, e13890.
    6. Rieske, P. et al. (2009) Exp. Cell Res. 315, 462.
    7. Ben-Zvi, A. et al. (2007) J. Neurosci. 27, 13000.
    8. Gil, C. et al. (2007) FEBS Lett. 581, 1851.
    9. Sandoval, M. et al. (2007) J. Neurochem. 101, 1672.
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