Anti-NMDAR2A (GluN2A) (extracellular) Antibody

NMDA receptor 2A, NR2A, GRIN2A, Ionotropic glutamate receptor subunit ε1, N-methyl-D-aspartate receptor subunit 2A
    Cat #: AGC-002
  • 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.
    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 0.55 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, ih, ip, lci, wb
    May also work in: ifc
    Western blot
    Western blot analysis of rat brain lysates:
    1. Anti-NMDAR2A (GluN2A) (extracellular) Antibody (#AGC-002), (1:600).
    2. Anti-NMDAR2A (GluN2A) (extracellular) Antibody, preincubated with the control peptide antigen.
    Immunoprecipitation
    Immunoprecipitation of rat brain lysates
    1. Cell lysate.
    2. Cell lysates + protein A beads + Anti-NMDAR2A (GluN2A) (extracellular) Antibody (#AGC-002).
    3. Cell lysates + protein A beads + pre-immune rabbit serum. 

    Black arrow indicates the NR2A protein while the red arrow shows the IgG heavy chain. Immunoblot was performed with Anti-NMDAR2A (GluN2A) (extracellular) Antibody.
    Immunohistochemistry
    Immuno-colocalization of GluN2A and GluN2B in mouse deep cerebellar nucleus
    Immunohistochemical staining of perfusion-fixed frozen mouse brain sections using Anti-NMDAR2B (GluN2B) (extracellular)-ATTO-594 Antibody (#AGC-003-AR), (1:60) and Anti-NMDAR2A (GluN2A) (extracellular) Antibody (#AGC-002), (1:200). A. Sections were incubated with Anti-NMDAR2A (GluN2A) (extracellular) Antibody, followed by goat anti-rabbit-Alexa-488 (green). B. The same sections were incubated with Anti-NMDAR2B (GluN2B) (extracellular)-ATTO-594 Antibody (red). C. Merge of A and B demonstrates the ubiquitous colocalization of the GluN2A and GluN2B subunits in cells with neuronal profiles in this nucleus. Arrows point at an example of NR2A and NR2B co-expression.
    Expression of NR2A in rat hippocampus
    Immunohistochemical staining of rat hippocampal dentate gyrus with Anti-NMDAR2A (GluN2A) (extracellular) Antibody (#AGC-002). A. NMDAR2A (green) appears diffusely in the outer molecular layer of the dentate gyrus (Out Mol.) and in cells along the subgranular layer (arrows). B. Staining of parvalbumin (PV, red) identifies interneurons in the dentate gyrus. C. Confocal merge demonstrates localization of PV in some neurons with NMDAR2A.
    Live cell imaging / Immunocytochemistry
    Expression of NR2A in rat C6 glioma cells
    Immunocytochemical staining of live intact rat C6 glioma cells. A. Cells were stained with Anti-NMDAR2A (GluN2A) (extracellular) Antibody (#AGC-002), (1:100), followed by goat anti-rabbit-AlexaFluor-555 secondary antibody (red). Cell nuclei were stained with the cell permeable dye Hoechst 33342 (blue staining). B. Live view of the same field.
    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.

    The 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) Antibody
    Expression of NR2A in human-induced pluripotent stem cell-derived neurons.Immunocytochemical staining of human-induced pluripotent stem cell-derived neurons using Anti-NMDAR2A (GluN2A) (extracellular) Antibody (#AGC-002). Adapted from Telezhkin, V. et al. (2016) Am. J. Physiol. 310, C250. with permission of The American Physiological Society.
    Last update: 15/11/2018

    Anti-NMDAR2A (GluN2A) (extracellular) Antibody (#AGC-002) 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. The antibody recognizes an extracellular epitope and is thus ideal for detecting GluN2A in living cells. It has been designed to recognize GluN2A from rat, mouse, and human samples.

    For research purposes only, not for human use
    Citations
    Western blot citations
    1. Mouse brain lysate (1:2000).
      Konstantoudaki, X. et al. (2016) Neuroscience 322, 333.
    2. Mouse hippocampus lysate (1:1000).
      Tindi, J.O. et al. (2015) J. Neurosci. 35, 8986.
    3. Mouse brain lysate (1:500).
      Atkin, G. et al. (2015) J. Neurosci. 35, 6165.
    Live cell imaging citations
    1. Rat hippocampal neurons.
      Dupuis, J.P. et al. (2014) EMBO J. 33, 842.
    2. Rat hippocampal primary neurons (1:500).
      Mikasova, L. et al. (2012) Brain 135, 1606.
    Immunohistochemistry citations
    1. Mouse brain sections (1:100).
      Atkin, G. et al. (2015) J. Neurosci. 35, 6165.
    Immunocytochemistry citations
    1. HEK293 transfected cells.
      Sibarov, D.A. et al. (2017) Front. Cell. Neurosci. 11, 155.
    2. Human induced pluripotent stem cells (1:300).
      Telezhkin, V. et al. (2016) Am. J. Physiol. 310, C250.
    3. Mouse hippocampal neurons.
      Atkin, G. et al. (2015) J. Neurosci. 35, 6165.
    4. Rat cultured hippocampal neurons (1:100).
      Swanger, S.A. et al. (2013) J. Neurosci. 33, 8898.
    More product citations
    1. Papouin, Tet al. (2012) Cell 150, 633.
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