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Anti-P2X7 Receptor (extracellular)-ATTO Fluor-488 Antibody

Purinergic receptor P2X7, P2RX7, P2Z, ATP receptor
Cat #: APR-008-AG
Alternative Name Purinergic receptor P2X7, P2RX7, P2Z, ATP receptor
  • KO Validated
  • Lyophilized Powder yes
    Type: Polyclonal
    Host: Rabbit
    Reactivity: h, m, r
    Immunogen
    • Peptide KKGWMDPQSKGIQTGRC, corresponding to amino acid residues 136-152 of mouse P2X7 receptor (Accession 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.
    RRID AB_2341045.
    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-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).
    Standard quality control of each lot Western blot analysis (unlabeled antibody, #APR-008), and direct flow cytometry (labeled antibody).
    Applications: fc, ic, if, ih, lci
    Immunohistochemistry
    • Mouse muscle-myenteric plexus sections (1:100) (Caputi, V. et al. (2017) Front. Pharmacol. 8, 350.).
    Direct flow cytometry
    • Cell surface detection of P2RX7 in intact living Jurkat cells:
      Cell surface detection of P2RX7 in intact living Jurkat cells:
      ___ Unstained cells.
      ___ Cells + Anti-P2X7 Receptor (extracellular)-ATTO Fluor-488 Antibody (#APR-008-AG), (10 µg/1x106 cells).
    Live cell imaging / Immunocytochemistry
    • Expression of P2RX7 in HEK-293 transfected cells
      Expression of P2RX7 in HEK-293 transfected cells
      Cell surface detection of P2RX7 in live intact P2X7-HEK-293 transfected cells. Extracellular staining of cells using Anti-P2X7 Receptor (extracellular)-ATTO Fluor-488 Antibody (#APR-008-AG), (green), (1:25).
    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 proinflamatory 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, IF- Immunofluorescence, 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)-ATTO Fluor-488 Antibody
    Expression of P2RX7 in rat colon.Immunohistochemical staining of rat distal colon section using Anti-P2X7 Receptor (extracellular)-ATTO Fluor-488 Antibody (#APR-008-AG). A. P2X7 staining (green) in normal colon sections co-localizes with that of HuC/D (red). B. P2X7 staining in the presence of colitis is detected in the neuromuscular layer and expression of the receptor is increased in myenteric ganglia. TOTO-3 (blue) was used to stain nuclei.Adapted from Antonioli, L. et al. (2014) PLoS One 9, e116253. with kind permission of Dr. Colucci, R. Div. Pharmacology and Chemotherapy, Dept. of Clinical and Experimental Medicine, University of Pisa Pisa, Italy.

    Last update: 23/08/2020

    Anti-P2X7 Receptor (extracellular) Antibody (#APR-008) is a highly specific antibody directed against an 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.

    Anti-P2X7 Receptor (extracellular)-ATTO Fluor-488 Antibody (#APR-008-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-P2X7 Receptor (extracellular)-ATTO Fluor-488 Antibody is specially suited to experiments requiring simultaneous labeling of different markers.

    For research purposes only, not for human use

    Applications

    Specifications

    Scientific Background

    Citations

    Citations
    Published figures using this product
    • Expression of P2X7 receptor in mouse longitudinal muscle-myenteric plexus sections.
      Expression of P2X7 receptor in mouse longitudinal muscle-myenteric plexus sections.
      Immunohistochemical staining of mouse longitudinal muscle-myenteric plexus sections using Anti-P2X7 Receptor (extracellular)-ATTO Fluor-488 Antibody (#APR-008-AG). P2X7 receptor staining (yellow) is detected in myenteric neurons.
      Adapted from Caputi, V. et al. (2017) Front. Pharmacol. 8, 350. with permission of Frontiers.
    KO validation citations
    1. Western blot analysis of mouse neutrophil lysate using #APR-008. Tested in P2X7-/- mice.
      Karmakar, M. et al. (2016) Nat. Commun. 7, 10555.
    Immunohistochemistry citations
    1. Mouse muscle-myenteric plexus sections (1:100).
      Caputi, V. et al. (2017) Front. Pharmacol. 8, 350.
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