Anti-P2X4 Receptor (extracellular) Antibody

P2RX4, P2X purinoceptor 4, ATP receptor
    Cat #: APR-024
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, m, r
    Immunogen
    Peptide (C)RDLAGKEQRTLTK, corresponding to amino acid residues 301-313 of rat P2X4 receptor (Accession P51577). Extracellular.
    Accession (Uniprot) Number P51577
    Gene ID 29659
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Mouse - 12/13 amino acid residues identical; human - 11/13 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.
    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 0.85 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, lci, wb
    May also work in: ifc, ih, ip
    Western blot
    Western blot analysis of rat brain membrane (lanes 1 and 5), mouse brain membrane (lanes 2 and 6), C6 cell line lysate (lanes 3 and 7) and SH-SY5Y cell line lysate (lanes 4 and 8):
    1-4. Anti-P2X4 Receptor (extracellular) Antibody (#APR-024), (1:200).
    2-8. Anti-P2X4 Receptor (extracellular) Antibody, preincubated with the control peptide antigen.
    Live cell imaging / Immunocytochemistry
    Expression of P2RX4 in rat pheochromocytoma cell line (PC12)
    Immunocytochemical staining of intact living PC12 cells. A. Extracellular staining of cells with Anti-P2X4 (extracellular) Antibody (#APR-024), (red, 1:50) followed by goat anti-rabbit-AlexaFluor-594 secondary antibody. B. Nuclear staining of cells using DAPI (blue). C. Merged images of A and B.
    References
    1. Prasad, M. et al. (2001) J. Physiol. 537, 667.
    2. Florenzano, F. et al. (2002) Neuroscience 115, 425.
    3. Ashcroft, F.M. et al. (2000) Ion Channels and Disease Ed 1, p. 405, Academic Press, San Diego.
    4. Khakh, B.S. et al. (2001) Pharmacol. Rev. 53, 107.
    5. Ding, Y. et al. (2000) J. Auton. Nerv. Syst. 81, 289.
    6. Le, K.T. et al. (1998) J. Neurosci. 18, 7152.
    7. Robertson, S.J. et al. (2001) Curr. Opin. Neurobiol. 11, 378.
    8. Dunn, P.M. et al. (2001) Prog. Neurobiol. 65, 107.
    9. Kim, M. et al. (2001) EMBO J. 20, 6347.
    10. Inoue, K. et al. (2004) J. Pharmacol. Sci. 94, 112.
    Scientific background

    The P2X purinergic receptors belong to the ligand-gated ion channel family and are activated by extracellular ATP.

    The structure and function of the P2X receptors, which were mainly investigated using in vitro models, indicate their involvement in synaptic communication, cell death, and differentiation.

    Seven mammalian P2X receptor subtypes (P2X1–P2X7) have been identified and cloned.1,2,3 All P2X receptor subtypes share the same structure of intracellular N- and C-termini, two membrane-spanning domains and a large extracellular loop.

    All P2X subunits can assemble to form homomeric or heteromeric functional channels with the exception of P2X6, which only appears to function as part of a heteromeric complex.4-9

    The various P2X receptors show distinct expression patterns. P2X1-6 have been found in the central and peripheral nervous system, while the P2X7 receptor is predominantly found in cells of the immune system.4

    The P2X2 receptor subunit has a widespread tissue distribution in autonomic neurons, but it is generally found to be co-expressed with one or more subtypes.

    Overexpression of P2X4 was demonstrated in microglia and in the spinal dorsal horn following peripheral nerve injury. It has been suggested that activation of P2X4 along with p38 MAPK is essential for the development of allodynia (pain from a stimulus that doesn't normally elicit pain) following nerve injury. Inhibition of P2X4 expression in spinal microglia has been suggested as a novel therapeutic approach for the treatment of allodynia.10

    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: 30/10/2018

    Anti-P2X4 Receptor (extracellular) Antibody (#APR-024) is a highly specific antibody directed against an epitope of the rat protein. The antibody can be used in western blot and immunocytochemistry applications. Anti-P2X4 Receptor (extracellular) Antibody recognizes an extracellular epitope and is thus ideal for detecting the receptor in living cells. It has been designed to recognize P2X4 purinergic receptor from mouse, human, and rat samples.

    For research purposes only, not for human use
    Citations
    Live cell imaging citations
    1. Mouse T-cells.
      Elkhal, A. et al. (2016) Sci. Rep. 6, 1.
    Related Products
      • Products for control experiments
        1. Anti-P2X4 Receptor Antibody (#APR-002). This antibody recognizes a different epitope and thus can also be used as a control for detecting P2X4 receptor.