Anti-p75 NGF Receptor (extracellular)-ATTO-550 Antibody

p75NTR, p75 Neurotrophin receptor, Low affinity NGF receptor, TNFR superfamily member 16, TNFRSF16, CD271
    Cat #: ANT-007-AO
    Alternative Name p75NTR, p75 Neurotrophin receptor, Low affinity NGF receptor, TNFR superfamily member 16, TNFRSF16, CD271
  • Lyophilized Powder
  • Antigen Incl.
  • Type: Polyclonal
    Host: Rabbit
    Reactivity: h, m, r
      • Peptide CEEIPGRWITRSTPPE corresponding to amino acid residues 188-203 of human p75NTR (Accession P08138). Extracellular, stalk domain.
        Anti-p75 NGF Receptor (extracellular)-ATTO-550 Antibody
    Accession (Uniprot) Number P08138
    Gene ID P08138
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Mouse - identical; rat - 15/16 amino acid residues identical.
    RRID AB_2039966.
    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-550. Maximum absorption 554 nm; maximum fluorescence 576 nm. The fluorescence is excited most efficiently in the 540 - 565 nm range. This label is related to the dye Rhodamine 6G and can be used with filters used to detect Rhodamine.
    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-007), and immunocytochemistry (labeled antibody).
    Applications: ic, if, ih, lci
      • Anti-p75 NGF Receptor (extracellular)-ATTO-550 Antibody
        Immuno-colocalization of LINGO-1 and p75NTR in rat hippocampus
        Immunohistochemical staining of immersion-fixed, free floating rat brain frozen sections using Anti-LINGO-1 (extracellular) Antibody (#ANT-032), (1:200) and Anti-p75 NGF Receptor (extracellular)-ATTO-550 Antibody (#ANT-007-AO), (1:80). A. LINGO-1 staining (green) in hippocampal CA3 region is detected in neuronal profiles (arrows). B. p75NTR staining in the same section (orange) is also seen in CA3 neurons (arrows). C. Merge of the two images demonstrates colocalization in several neurons (arrows). Cell nuclei are stained with DAPI (blue).
      • Anti-p75 NGF Receptor (extracellular)-ATTO-550 Antibody
        Expression of p75NTR in p75NTR-transfected 3T3 cells
        A. Cell surface detection of p75NTR in intact live p75NTR-transfected 3T3 cells with Anti-p75 NGF Receptor (extracellular)-ATTO-550 Antibody (#ANT-007-AO), (1:100). B. Live view of the same field as in A.
        Anti-p75 NGF Receptor (extracellular)-ATTO-550 Antibody
        Immuno-colocalization of p75NTR and mGluR1 in rat DRGs
        Cell surface detection of p75NTR and mGluR1 in intact live DRG neurons labeled with Anti-mGluR1 (extracellular) Antibody (#AGC-006), (1:100) followed by goat-anti-rabbit-AlexaFluor-488 secondary antibody (green). The cells were then labeled with Anti-p75 NGF Receptor (extracellular)-ATTO-550 Antibody (#ANT-007-AO), (1:50), (red). Nuclei were visualized using the cell-permeable DNA binding dye Hoechst 33342 (blue). Note that the Anti-p75 NGF Receptor (extracellular)-ATTO-550 Antibody does not stain all DRG neurons as expected. Colocalization is observed in some nerve fibers of the mGluR1 and p75NTR receptors.
    References
      • 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,
    IF- Immunofluorescence, IHC- Immunohistochemistry, IP- Immunoprecipitation, LCI- Live cell imaging, N- Neutralization, WB- Western blot

    Species reactivity key:

    H- Human, M- Mouse, R- Rat
    Image & Title:


    Expression and visualization of p75NTR in live intact dorsal root ganglion (DRG) explants DRG intact explants were incubated with Anti-p75 NGF Receptor (extracellular)-ATTO-550 Antibody (#ANT-007-AO), (1:50), under conditions that allowed receptor internalization. Retrograde transport of the internalized receptor (orange) was visualized in vesicles moving across the axon. Right side is distal axon while left is explant side. Frames were taken every 2 seconds. Movie is 10 frames per second.
    Movie was kindly provided by Dr. Eran Perlsson, the Dept. of Pharmacology and Physiology, Tel-Aviv University.

    Last update: 07/11/2019

    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-550 Antibody (#ANT-007-AO) is directly labeled with an ATTO-550 fluorescent dye. ATTO dyes are characterized by strong absorption (high extinction coefficient), high fluorescence quantum yield, and high photo-stability. The ATTO-550 fluorescent label is related to the well known dye Rhodamine 6G and can be used with filters typically used to detect Rhodamine. Anti-p75 NGF Receptor (extracellular)-ATTO-550 Antibody is especially suited for experiments requiring simultaneous labeling of different markers.

    For research purposes only, not for human use
    Citations
      • Anti-p75 NGF Receptor (extracellular)-ATTO-550 Antibody
        Co-localization of RABV with p75NTR in DRG neuron tips.
        Immunocytochemical staining of mouse DRGs using TIRF imaging. RABV-EGFP and Anti-p75 NGF Receptor (extracellular)-ATTO-550 Antibody (#ANT-007-AO) show that the virus and p75NTR particles shift from the periphery to the center of the growth cone, where they are internalized into the cell. Lower panel show co-localized puncta (left) shifting towards the center of the growth cone (middle) until internalized (right).
        Adapted from Gluska, S. et al. (2014) with kind permission from Prof. Eran Perlson, Dpt. Physiology and Pharmacology, Sackler Faculty of Medicine, Tel Aviv University, Israel.
      • mouse DRGs.
        Gluska, S. et al. (2014) PLoS Pathog. 10, e1004348.
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