Anti-NMDAR2A (GluN2A) (extracellular)-ATTO-488 Antibody

NMDA receptor 2A, NR2A, GRIN2A, Ionotropic glutamate receptor subunit ε1, N-methyl-D-aspartate receptor subunit 2A
    Cat #: AGC-002-AG
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, m, r
    Immunogen
    Peptide GHSHDVTERELRN(C), corresponding to amino acid residues 41-53 of rat NR2A (Accession Q00959). Extracellular, N-terminus.
    Accession (Uniprot) Number Q00959
    Gene ID 24409
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Human, mouse - 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.
    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).
    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 (unlabeled antibody,#AGC-002), and immunohistochemistry (labeled antibody).
    Applications: ih
    May also work in: ic, lci
    Immunohistochemistry
    Expression of NR2A in mouse brain sections
    Immunohistochemical staining of perfusion-fixed frozen mouse brain sections using Anti-NMDAR2A (GluN2A) (extracellular)-ATTO-488 Antibody (#AGC-002-AG), (1:80). A. NR2A staining (green) in striatum is detected in neuronal profiles. B. NR2A staining in cingulate cortex is also detected in neuronal profiles. Cell nuclei are labeled with DAPI (blue).
    References
    1. Dingledine, R. et al. (1999) Pharmacol. Rev. 51, 7.
    2. Mayer, M.L. and Armstrong, N. (2004) Annu. Rev. Physiol66, 161.
    3. Prybylowski, K. and Wenthold, R.J. (2004) J. Biol. Chem279, 9673.
    4. Mayer, M.L. (2006) Nature 440, 456.
    Scientific background

    The NMDA receptors are members of the glutamate receptor family of ion channels that also include the AMPA and Kainate receptors.

    The NMDA receptors are encoded by seven genes: one NMDAR1 (or NR1) subunit, four NR2 (NR2A-NR2D) and two NR3 (NR3A-NR3B) subunits. The functional NMDA receptor appears to be a heterotetramer composed of two NMDAR1 and two NMDAR2 subunits. Whereas the NMDAR2 subunits that assemble with the NMDAR1 subunit can be either of the same kind (i.e. two NMDAR2A subunits) or different (one NMDAR2A with one NMDAR2B). NMDAR3 subunits can substitute the NMDAR2 subunits in their complex with the NMDAR1 subunit.

    NMDAR is unique among ligand-gated ion channels in that it requires the simultaneous binding of two obligatory agonists: glycine and glutamate that bind to the NMDAR1 and NMDAR2 binding sites respectively. Another unique characteristic of the NMDA receptors is their dependence on membrane potential. At resting membrane potentials the channels are blocked by extracellular Mg2+. Neuronal depolarization relieves the Mg2+ blockage and allows ion influx into the cells. NMDA receptors are strongly selective for Ca2+ influx differing from the other glutamate receptor ion channels that are non-selective cation channels.

    Ca2+ entry through the NMDAR regulates numerous downstream signaling pathways including long term potentiation (a molecular model of memory) and synaptic plasticity that may underlie learning. In addition, the NMDA receptors have been implicated in a variety of neurological disorders including epilepsy, ischemic brain damage, Parkinson’s and Alzheimer’s disease.

    NMDA receptors expression and function are modulated by a variety of factors including receptor trafficking to the synapses and internalization as well as phosphorylation and interaction with other intracellular proteins.

    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-NMDAR2A (GluN2A) (extracellular)-ATTO-488 AntibodyImmuno-colocalization of GluN2A and SynGAP in rat cingulate cortex.Immunohistochemical staining of immersion-fixed, free floating rat brain frozen sections using Anti-NMDAR2A (GluN2A) (extracellular)-ATTO-488 Antibody (#AGC-002-AG), (1:200) and Anti-SynGAP Antibody (#APZ-032), (1:200), followed by donkey-anti-rabbit-Cy3. A. GluN2A staining (green) appears in neuronal profiles (arrows). B. SynGAP staining (red) is detected mostly in apical dendrites (horizontal arrows) and in neuronal soma (vertical arrows). C. Merge of the two images demonstrates colocalization in some neurons (vertical arrows). Cell nuclei are stained with DAPI (blue).

    Last update: 15/11/2018

    Anti-NMDAR2A (GluN2A) (extracellular) Antibody (#AGC-002) is a highly specific antibody directed against an extracellular epitope of the rat protein. The antibody can be used in western blot, immunoprecipitation, immunocytochemistry, and immunohistochemistry. It has been designed to recognize GluN2A from rat, mouse, and human samples.

    Anti-NMDAR2A (GluN2A) (extracellular)-ATTO-488 Antibody (#AGC-002-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-NMDAR2A (GluN2A) (extracellular)-ATTO-488 Antibody has been tested in immunohistochemistry and is specially suited to experiments requiring simultaneous labeling of different markers.

    For research purposes only, not for human use