Anti-P2X7 Receptor Antibody

Purinergic receptor P2X7, P2RX7, P2Z, ATP receptor
Cat #: APR-004
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, m, r
    Immunogen
    Peptide (C)KIRKEFPKTQGQYSGFKYPY, corresponding to amino acid residues 576-595 of rat P2X7 receptor (Accession Q64663). Intracellular, C-terminus.
    Accession (Uniprot) Number Q64663
    Gene ID 29665
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Mouse - 18/20 amino acid residues identical; human - 16/20 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 0.6 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, wb
    Western blot
    Western blot analysis of rat hippocampus lysates:
    1. Anti-P2X7 Receptor Antibody (#APR-004), (1:200).
    2. Anti-P2X7 Receptor Antibody, preincubated with the control peptide antigen.
    Immunoprecipitation
    Mouse brain, lung and submandibular glands (Sim, J.A. et al. (2004) J. Neurosci. 24, 6307.).
    Immunohistochemistry
    Expression of P2X7 Receptor in rat pancreas
    Immunohistochemical staining of rat paraffin-embedded endocrine and exocrine pancreas sections using Anti-P2X7 Receptor Antibody (#APR-004), (1:50), followed by goat anti-rabbit-AlexaFluor-555-secondary antibody (1:500), (red). Staining is highly specific for endocrine cells of the Isle of Langerhans. Hoechst 33342 is used as the counterstain (blue).
    Immunocytochemistry
    Mouse pancreatic stellate cells (PSCs), (1:100), (Haanes, K.A. et al. (2012) PLoS ONE 7, e51164.).

    Human MG63 and SaOS2 osteosarcoma cell lines (Alqallaf, S.M. et al. (2009) Br. J. Pharmacol. 156, 1124.).
    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.
    Scientific background

    The P2X7 purinergic receptor is a member of the ionotropic P2X receptor family that is activated by ATP. To date, this family consists of seven receptor subtypes, named P2X1-P2X7, all of which have been cloned.

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

    The P2X7 receptor mediates the release of proinflamatory cytokines and stimulation of transcription factors and may also play an important role in apoptosis.1-3

    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 Antibody
    Knockout validation of Anti-P2X7 Receptor Antibody in mouse spinal cord.
    Western blot analysis of mouse total lumbar spinal cord lysates (L3–L5) from WT, P2X7+/- and P2X7-/- 90-day-old mice using Anti-P2X7 Receptor Antibody (#APR-004).Adapted from Apolloni, S. et al. (2013) Hum. Mol. Genet. 22, 4102. with permission of Oxford University Press.

    Last update: 14/03/2019

    Anti-P2X7 Receptor Antibody (#APR-004) is a highly specific antibody directed against an epitope of the rat protein. The antibody can be used in western blot, immunoprecipitation, immunocytochemistry, and immunohistochemistry applications. It has been designed to recognize P2X7 purinergic receptor from rat, mouse, and human samples.

    For research purposes only, not for human use
    Citations
    Published figures using this product
    Expression 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.
    KO validation citations
    1. Western blot analysis of mouse neutrophil lysate. Also tested on P2X7-/- mice.
      Karmakar, M. et al. (2016) Nat. Commun. 7, 10555.
    2. Western blot analysis of mouse GL261 glioma cell lysate. Also tested on siRNA treated cells.
      Gehring, M.P. et al. (2015) Int. J. Biochem. Cell Biol. 68, 92.
    3. Immunocytochemical staining of mouse microglial cells (1:600). Also tested in P2X7-/- cells.
      Fischer, W. et al. (2014) Purinergic Signal. 10, 313.
    4. Western blot analysis of mouse lumbar spinal cord protein lysates (1:500). Also tested in P2X7-/- mice.
      Apolloni, S. et al. (2013) Hum. Mol. Genet. 22, 4102.
    Western blot citations
    1. Mouse MC3T3-E1 osteoblastic cells.
      Zhang, Q. et al. (2016) Biochem. Biophys. Res. Commun. 476, 438.
    2. Mouse bone lysate and osteoblast MOB-C cell lysate.
      Seref-Ferlengez, Z. et al. (2016) PLoS ONE 11, e0155107.
    3. HEK 293 transfected cells.
      Aprile-Garcia, F. et al. (2016) PLoS ONE 11, e0151862.
    4. Mouse and human brain lysates.
      Jimenez-Pacheco, A. et al. (2016) J. Neurosci. 36, 5920.
    5. Mouse neutrophil lysate. Also tested on P2X7-/- mice.
      Karmakar, M. et al. (2016) Nat. Commun. 7, 10555.
    6. Rat brain lysate (1:200).
      Grygorowizc, T. et al. (2016) Mol. Cell. Neurosci. 74, 1.
    7. Mouse GL261 glioma cell lysate. Also tested on siRNA treated cells.
      Gehring, M.P. et al. (2015) Int. J. Biochem. Cell Biol. 68, 92.
    8. Rat cerebral cortex synaptosomal membranes (1:300).
      Barros-Barbosa, A.R. et al. (2015) Neuroscience 306, 74.
    9. HEK293-TSA 201 transfected cell lysates (1:5000).
      Allsopp, R. and Evans, R.J. (2015) J. Biol. Chem. 290, 14556.
    10. Mouse lumbar spinal cord protein lysates (1:500). Also tested in P2X7-/- mice.
      Apolloni, S. et al. (2013) Hum. Mol. Genet. 22, 4102.
    11. Rat Schwann cell lysate (1:500).
      Faroni, A. et al. (2013) Cell Death Dis. 4, e743.
    12. Mouse and human cortex lysates.
      Jimenez Pacheco, A. et al. (2013) Epilepsia 54, 1551.
    13. Rat eye lysate (1:500).
      Sugiyama, T. et al. (2013) Mol. Vis 19, 2080.
    14. Rat hippocampus lysate (1:1000).
      Yu, Q. et al. (2013) Purinergic Signal. 9, 663.
    15. Mouse astrocyte culture lysate (1:500).
      Okuda, H. et al. (2013) Life Sci. 92, 808.
    16. Rat PC12 cells (1:1000).
      Xu, H. et al. (2013) Brain Res. Bull. 94, 63.
    17. Mouse cerebral synaptosomes.
      Naviaux, R.K. et al. (2013) PLoS ONE 8, e57380.
    18. Rat L4-5 spinal cord lysates (1:2000).
      Kobayashi, K. et al. (2011) Neurosci. Lett. 504, 57.
    19. Mouse microgial cells (1:200).
      Witting, A. et al. (2004) Proc. Natl. Acad. Sci. U.S.A. 101, 3214.
    Immunoprecipitation citations
    1. Mouse bone lysate.
      Seref-Ferlengez, Z. et al. (2016) PLoS ONE 11, e0155107.
    2. Mouse brain, lung and submandibular glands.
      Sim, J.A. et al. (2004) J. Neurosci. 24, 6307.
    Immunohistochemistry citations
    1. Rat brain sections (1:50).
      Barros-Barbosa, A.R. et al. (2015) Neuroscience 306, 74.
    2. Rat spinal cord sections.
      Vazquez-Villoldo, N. et al. (2014) Glia 62, 171.
    3. Rat brain sections (1:1000).
      Yu, Q. et al. (2013) Purinergic Signal. 9, 663.
    4. Rat retina sections (1:200).
      Sugiyama, T. et al. (2013) Mol. Vis 19, 2080.
    Immunocytochemistry citations
    1. Rat synaptosomes (1:50).
      Barros-Barbosa, A.R. et al. (2015) Neuroscience 306, 74.
    2. Mouse microglial cells (1:600). Also tested in P2X7-/- cells.
      Fischer, W. et al. (2014) Purinergic Signal. 10, 313.
    3. Human ESC NPs (1:100).
      Forostyak, O. et al. (2013) Stem Cells Dev. 22, 1506.
    4. Rat Schwann cells (1:1000).
      Faroni, A. et al. (2013) Cell Death Dis. 4, e743.
    5. Rat PC12 cells.
      Xu, H. et al. (2013) Brain Res. Bull. 94, 63.
    6. Mouse pancreatic stellate cells (PSCs), (1:100).
      Haanes, K.A. et al. (2012) PLoS ONE 7, e51164.
    7. Human MG63 and SaOS2 osteosarcoma cell lines.
      Alqallaf, S.M. et al. (2009) Br. J. Pharmacol. 156, 1124.
    Indirect flow cytometry citations
    1. Mouse peritoneal macrophage.
      Zanin, R.F. et al. (2015) Purinergic Signal. 11, 463.
    More product citations
    1. Brown, L.E. et al. (2013) J. Neurosci. 33, 3557.
    2. Deplano, S. et al. (2013) J. Leukoc. Biol. 93, 127.
    3. Kakurai, K. et al. (2013) Neurosci. Lett. 534, 237.
    4. Murasaki, K. et al. (2013) J. Dental Res. 92, 260.
    5. Traves, P.G. et al. (2013) J. Immunol. 190, 4226.
    6. Garcia-Huerta, P. et al. (2012) J. Biol. Chem. 287, 44628.
    7. Le Gall, S.M. et al. (2012) PLoS ONE 7, e52161.
    8. Kim, M. et al. (2001) J. Biol. Chem. 276, 23262.
    9. Kim, M. et al. (2001) EMBO J. 20, 6347.
    10. Mehta, V.B. et al. (2001) J. Biol. Chem. 276, 3820.
    11. Solle, M. et al. (2001) J. Biol. Chem. 276, 125.
    12. Gitterman, D.P. and Evans, R.J. (2000) Br. J. Pharmacol. 131, 1561.
    13. Lewis, C.J. and Evans, R.J. (2000) Br. J. Pharmacol. 131, 1659.
    14. Koshlukova, S.E. et al. (2000) Infect. Immun. 68, 6848.
    15. Vial, C. and Evans, R.J. (2000) Br. J. Pharmacol. 131, 1489.
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