Anti-p75 NGF Receptor (extracellular)-ATTO-488 Antibody

p75NTR, p75 Neurotrophin receptor, Low affinity NGF receptor, TNFR superfamily member 16, TNFRSF16, CD271
    Cat #: ANT-007-AG
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, m, r
    Immunogen
    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.
    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).
    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 (unlabeled antibody, #ANT-007), and immunocytochemistry (labeled antibody).
    Applications: fc, ic, ih, lci
    Immunohistochemistry
    Immunolocalization of p75NTR and TrkA in rat dorsal root ganglion
    Immunohistochemical staining of perfusion-fixed frozen rat dorsal root ganglia (DRG) sections using Anti-p75 NGF Receptor (extracellular)-ATTO-488 Antibody (#ANT-007-AG), (1:60) and Anti-TrkA (extracellular)-ATTO-633 Antibody (#ANT-018-FR), (1:60). A. Sections were labeled with Anti-p75 NGF Receptor (extracellular)-ATTO-488 Antibody (green). B. The same section was stained with Anti-TrkA (extracellular)-ATTO-633 Antibody (purple). C. Merge of the two images suggests extensive co-localization (arrow points at an example) of TrkA and p75NTR receptors in rat dorsal root ganglion. Cell nuclei were stained with DAPI (blue).
    Direct flow cytometry
    Direct flow cytometry analysis of live intact rat pheochromocytoma (PC12) cells:
    ___ Unstained cells.
    ___ Cells + Anti-p75 NGF Receptor (extracellular)-ATTO-488 Antibody (#ANT-007-AG), (1:20).
    Live cell imaging / Immunocytochemistry
    Expression of p75NTR in human A875 melanoma cells
    Immunocytochemical staining of live intact human A875 melanoma cells. A. Cells were stained with Anti-p75 NGF Receptor (extracellular)-ATTO-488 Antibody (#ANT-007-AG), (1:50), (green). B. Live image of the cells. C. Merge of the two images.
    References
    1. Hempstead, B.L. (2002) Curr. Opin. Neurobiol. 12, 260.
    2. Chao, M.V. (2003) Nat. Rev. Neurosci4, 299.
    3. Barker, P.A. (2004) Neuron 42, 529.
    4. Dechant, G. and Barde, Y.A. (2002) Nat. Neurosci. 5, 1131.
    Scientific background

    The p75 neurotrophin receptor (p75NTR) is a member of the tumor necrosis factor (TNF) receptor superfamily. Like all members of this superfamily, p75NTR is a type I transmembrane protein with tandem cysteine-rich domains in its extracellular portion, as well as an intracellular death domain. p75NTR binds to its ligands as a homodimer but can also form heterodimers with other receptors such as TrkA, TrkB, TrkC, Nogo receptor, and sortilin. The precise multimeric receptor complex formed between p75NTR and the other receptors determine the ligand being recognized (see below) and the biological response to its binding1-3.

    As its name implies, p75NTR binds to all the neurotrophins (NTR) (NGF, BDNF, NT-3, and NT-4) with similar nanomolar affinities. Co-expression of p75NTR with the Trk receptors enhances its 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, leading to cell death by apoptosis1-3. Finally, a multimeric complex of p75NTR and Nogo receptor binds to myelin proteins such as Nogo-66, MAG, and OmgP, resulting in inhibition of axonal growth4.

    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: 29/10/2018

    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.

    Anti-p75 NGF Receptor (extracellular)-ATTO-488 Antibody (#ANT-007-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-p75 NGF Receptor (extracellular)-ATTO-488 Antibody is specially suited to experiments requiring simultaneous labeling of different markers.

    For research purposes only, not for human use
    Citations
    Related Products