Anti-P2X7 Receptor (extracellular) Antibody

Purinergic receptor P2X7, P2RX7, P2Z, ATP receptor
    Cat #: APR-008
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, m, r
    Immunogen
    Peptide KKGWMDPQSKGIQTGRC, corresponding to amino acid residues 136-152 of mouse P2X7 receptor (Accession number Q9Z1M0). Extracellular loop.
    Accession (Uniprot) Number Q9Z1M0
    Gene ID 18439
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Human, rat - identical; bovine - 14/17 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 0.8 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 1,5) and human cell lines; K562 (lanes 2 and 6); WEHI-231 (lanes 3 and 7) and HL-60 (lanes 4 and 8): 
    1-4. Anti-P2X7 Receptor (extracellular) Antibody (#APR-008), (1:200).  
    5-8. Anti-P2X7 Receptor (extracellular) Antibody, preincubated with the control peptide antigen.
    Immunohistochemistry
    Expression of P2RX7 in rat pancreas
    Immunohistochemical staining of rat pancreas  frozen section using Anti-P2X7 Receptor (extracellular) Antibody (#APR-008), (1:100), followed by mouse anti-rabbit-AlexaFluor-594 secondary antibody (red). Staining is present in endocrine cells of the Isle of Langerhans. Hoechst 33342 is used as the counterstain.
    Immunocytochemistry
    Mouse P2X7 transfected in CHO cells (1:500). (Adriouch, S. et al. (2005) Cell. Immunol236, 72.).
    Indirect flow cytometry
    5-10 µg antibody/1x106 cells.
    The control antigen is not suitable for this application.
    Live cell imaging / Immunocytochemistry
    Expression of P2RX7 in rat RBL cells
    Immunocytochemical staining of intact living rat basophilic leukemia (RBL) cells. A. Extracellular staining of cells using Anti-P2X7 Receptor (extracellular) Antibody (#APR-008), (1:100) followed by goat anti-rabbit-AlexaFluor-594 secondary antibody (red). B. Live view of the cells.
    References
    1. Ding, Y. et al. (2000) J. Auton. Nerv. Syst. 81, 289.
    2. Kim, M. et al. (2001) EMBO J. 20, 6347.
    3. Chizh, B.A. and Illes, P. (2001) Pharmacol. Rev. 53, 553.
    4. Nihei, O.K. et al. (2000) Mem.Inst.Oswaldo Cruz. 95, 415.
    Scientific background

    The P2X7 receptor is a member of the ionotropic P2X receptor family that is activated by ATP. To date, this family is composed of seven cloned receptor subtypes, named P2X1-P2X7.  

    The different P2X receptors show distinct expression patterns. P2X1-6 have been found in the central and peripheral nervous system, while the P2X7 receptor is found in cells of the immune system, particularly antigen presenting cells, and microglia. The P2X7 receptor mediates the release of proinflammatory cytokines, stimulation of transcription factors and may also have an important role in apoptosis.1-3

    Different techniques have been used to characterize the P2X7 receptor. Most of them investigated pores, ion channels (electrophysiology) and membrane alterations (calcium microfluorometry, dye uptake, membrane depolarization and ion influx analysis). With the introduction of flow cytometry, it is now possible to analyze multiple cell parameters such as cell cycle, cell membrane alteration, calcium influx and cell phenotype.4

    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
    Image & Title:

    Anti-P2X7 Receptor (extracellular) AntibodyExpression and upregulation of microglial P2X7 receptor in cancer.A. Immunohistochemical staining of of rat spinal dorsal horn using Anti-P2X7 Antibody (#APR-004) or Anti-P2X7 (extracellular) Antibody (#APR-008) shows that P2X7 co-localizes with CD11b, a microglia marker (two panels) and not with NeuN, a neuronal marker (lower right panel). Specificity of the antibody was tested on P2X7 knock-out mice (western blot, in upper panel). B. Western blot analysis of rat dorsal horn lysates shows that P2X7 expression increases on post-tumor day 14 (PTD 14).Adapted from Yang, Y. et al. (2015) with permission of the Society for Neuroscience.

    Last update: 20/11/2018

    Anti-P2X7 Receptor (extracellular) Antibody (#APR-008) is a highly specific antibody directed against an extracellular epitope of the mouse protein. The antibody can be used in western blot, indirect flow cytometry, immunohistochemistry, live cell imaging, and immunocytochemistry applications. It has been designed to recognize P2X7 purinergic receptor from mouse, rat, and human samples.

    For research purposes only, not for human use
    Citations
    KO validation citations
    1. Western blot analysis on mouse neutrophil lysate. Also tested on P2X7-/- mice.
      Karmakar, M. et al. (2016) Nat. Commun. 7, 10555.
    Western blot citations
    1. HEK 293 transfected cells (1:10000).
      Migita, K. et al. (2016) Mol. Pharmacol. 90, 116.
    2. Mouse neutrophil lysate. Also tested on P2X7-/- mice.
      Karmakar, M. et al. (2016) Nat. Commun. 7, 10555.
    3. Mouse MC3T3-E1 cells lysate (preosteoblast-like cell line).
      Gangadharan, V. et al. (2015) Am. J. Physiol. 308, C41.
    4. Rat eye lysate (1:500).
      Sugiyama, T. et al. (2013) Mol. Vis 19, 2080.
    5. Human immortalized gingival keratinocyte (HIGK) cell lysate.
      Hung, S.C. et al. (2013) PLoS ONE 8, e70210.
    6. Equine hoof lysate (1:200).
      Zamboulis, D.E. et al. (2013) Purinergic Signal. 9, 383.
    7. Human mesenchymal stem cells (hMSCs).
      Sun, D. et al. (2013) Stem Cells 31, 1170.
    Live cell imaging citations
    1. Mouse T-cells.
      Elkhal, A. et al. (2016) Sci. Rep. 6, 1.
    Immunohistochemistry citations
    1. Mouse retinal sections.
      Hu, S.J. et al. (2015) J. Neurosci. 35, 6987.
    2. Rat retina sections (1:200).
      Sugiyama, T. et al. (2013) Mol. Vis 19, 2080.
    3. Equine palmar digital artery and Schwann cells (1:200).
      Zamboulis, D.E. et al. (2013) Purinergic Signal. 9, 383.
    4. Primate retina (1:500).
      Gu, B.J. et al. (2013) FASEB J. 27, 1479.
    Immunocytochemistry citations
    1. HEK 293 transfected cells (1:1000).
      Migita, K. et al. (2016) Mol. Pharmacol. 90, 116.
    2. Human mesenchymal stem cells (hMSCs) (1:100).
      Sun, D. et al. (2013) Stem Cells 31, 1170.
    3. Mouse P2X7 transfected in CHO cells (1:500).
      Adriouch, S. et al. (2005) Cell. Immunol236, 72.
    Neutralization citations
    1. Antibody was used to block P2X7-dependent PGE2 release.
      Li, J. et al. (2005) J. Biol. Chem. 280, 42952.
    More product citations
    1. Takai, E. et al. (2012) J. Cell Sci. 125, 5051.
    Related Products
      • Products for control experiments
        1. Anti-P2X7 Receptor Antibody (#APR-004). This antibody recognizes a different epitope and thus can also be used as a control for detecting P2X7.