Anti-NMDAR2B (GluN2B) (extracellular) Antibody

NMDA receptor 2B, Ionotropic glutamate receptor subunit ε2, N-methyl-D-aspartate receptor subunit 2B, GRIN2B, NR2B
    Cat #: AGC-003
    Alternative Name NMDA receptor 2B, Ionotropic glutamate receptor subunit ε2, N-methyl-D-aspartate receptor subunit 2B, GRIN2B, NR2B
  • Lyophilized Powder
  • Antigen Incl.
  • Type: Polyclonal
    Host: Rabbit
    Reactivity: h, m, r
    Immunogen
    • Peptide (C)NTHEKRIYQSNMLNR, corresponding to amino acid residues 323-337 of rat NMDA receptor 2B (Accession Q00960). Extracellular, N-terminus.
    • Anti-NMDAR2B (GluN2B) (extracellular) Antibody
    Accession (Uniprot) Number Q00960
    Gene ID 24410
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Human, mouse - identical.
    RRID AB_2040028.
    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.
    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).
    Negative 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.
    Negative control antigen reconstitution 100 µl double distilled water (DDW).
    Negative 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, if, ih, ip, lci, wb
    May also work in: ifc*
    Western blot
    • Anti-NMDAR2B (GluN2B) (extracellular) Antibody
      Western blot analysis of rat brain lysates:
      1. Anti-NMDAR2B (GluN2B) (extracellular) Antibody (#AGC-003), (1:600).
      2. Anti-NMDAR2B (GluN2B) (extracellular) Antibody, preincubated with the negative control antigen.
    • Mouse forebrain cortex (1:200) (Fontaine, R.H. et al. (2008) Proc. Natl. Acad. Sci. U.S.A. 105, 16779.).
    Immunoprecipitation
    • Anti-NMDAR2B (GluN2B) (extracellular) Antibody
      Immunoprecipitation of rat brain lysates:
      1. Cell lysates.
      2. Cell lysates + protein A beads + Anti-NMDAR2B (GluN2B) (extracellular) Antibody (#AGC-003). 
      3. Cell lysates + protein A beads + pre-immune rabbit serum.  

      Red arrow indicates the NMDA receptor 2B protein while the black arrow shows the IgG heavy chain. Immunoblot was performed with Anti-NMDAR2B (GluN2B) (extracellular) Antibody.
    Immunohistochemistry
    • Anti-NMDAR2B (GluN2B) (extracellular) Antibody
      Expression of NMDA receptor 2B in rat hippocampus
      Immunohistochemical staining of rat hippocampal CA1 frozen sections stained with Anti-NMDAR2B (GluN2B) (extracellular) Antibody (#AGC-003), (1:100). A. NMDAR2B (green) appears in the pyramidal layer of CA1. B. Staining of neurofilament 200 (red) identifies neuronal processes. C. Confocal merge demonstrates localization of NMDAR2B in cells and not in processes.
    • Anti-NMDAR2B (GluN2B) (extracellular) Antibody
      Expression of NMDA receptor 2B in rat cortex
      Immunohistochemical staining of rat parietal cortex frozen sections stained with Anti-NMDAR2B (GluN2B) (extracellular) Antibody (#AGC-003), (1:100). A. NMDAR2B (green) appears in the pyramidal layer of layer 5. B. Staining of neurofilament 200 (red) identifies neuronal processes. C. Confocal merge demonstrates localization of NMDAR2B in cells.
    Immunocytochemistry
    • Mouse striatal cells (Del Toro, D. et al. (2010) J. Neurochem. 115, 153.).
    Live cell imaging / Immunocytochemistry
    • Rat hippocampal primary neurons (1:500) (Mikasova, L. et al. (2012) Brain 135, 1606.).
    References
    1. Dingledine, R. et al. (1999) Pharmacol. Rev. 51, 7.
    2. Mayer, M.L. and Armstrong, N. (2004) Annu. Rev. Physiol. 66, 161.
    3. Prybylowski, K. and Wenthold, R.J. (2004) J. Biol. Chem. 279, 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, 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-NMDAR2B (GluN2B) (extracellular) AntibodyExpression of GluN2B in human-induced pluripotent stem cell-derived neurons.Immunocytochemical staining of human-induced pluripotent stem cell-derived neurons using Anti-NMDAR2B (GluN2B) (extracellular) Antibody (#AGC-003).Adapted from Telezhkin, V. et al. (2016) Am. J. Physiol. 310, C250. with permission of The American Physiological Society.

    Last update: 03/03/2020

    Anti-NMDAR2B (GluN2B) (extracellular) Antibody (#AGC-003) is a highly specific antibody directed against an epitope of the rat protein. The antibody can be used in western blot, immunocytochemistry, live cell imaging, immunohistochemistry, and immunoprecipitation applications. It has been designed to recognize GluN2B from human, rat, and mouse samples.

    For research purposes only, not for human use

    Applications

    Specifications

    Scientific Background

    Citations

    Citations
    Published figures using this product
    • Anti-NMDAR2B (GluN2B) (extracellular) Antibody
      Expression of GluN2B in rat hippocampal neurons
      Immunocytochemistry of living rat dissociated hippocampal neurons. Extracellular staining of cell with Anti-NMDAR2B (GluN2B) (extracellular) Antibody (#AGC-003). GluN2B cell surface expression (green) increases following extracellular matrix removal (lower panels). GluN2B expression coincides with PSD-95 synaptic marker.
      Adapted from Schweitzer, B. et al. (2017) Sci. Rep. 7, 10991. with permission of SPRINGER NATURE.
    Western blot citations
    1. Mouse striatal tissue lysate.
      Kang, R. et al. (2019) Front. Synaptic Neurosci. 11, 3.
    2. Rat primary hippocampal neuron lysate.
      Schweitzer, B. et al. (2017) Sci. Rep. 7, 10991.
    3. Mouse cortex lysate.
      Lin, H. et al. (2016) Front. Cell. Neurosci. 10, 34.
    4. Mouse brain lysate.
      Wang, Y.C. et al. (2016) J. Cereb. Blood Flow Metab. 37, 980.
    5. Mouse hippocampus lysate.
      Antonelli, R. et al. (2016) J. Neurosci. 36, 5437.
    6. Mouse brain lysate (1:2000).
      Konstantoudaki, X. et al. (2016) Neuroscience 322, 333.
    7. Mouse hippocampus lysate (1:1000).
      Tindi, J.O. et al. (2015) J. Neurosci. 35, 8986.
    8. Mouse brain lysate (1:500).
      Atkin, G. et al. (2015) J. Neurosci. 35, 6165.
    9. Mouse striatal lysate (1:500).
      Dau, A. et al. (2014) Neurobiol. Dis. 62, 533.
    Live cell imaging citations
    1. Mouse embryonic hippocampal neurons.
      Morini, R. et al. (2018) Front. Mol. Neurosci. 11, 313.
    2. Mouse dissociated hippocampal culture.
      Altmuller, F. et al. (2017) PLoS Genet. 13, e1006684.
    3. Rat hippocampal culture.
      Ferreira, J.S. et al. (2017) eLife 6, 8986.
    4. Rat hippocampal culture.
      Joshi, P. et al. (2017) Sci. Rep. 7, 41734.
    5. Rat primary hippocampal neurons.
      Schweitzer, B. et al. (2017) Sci. Rep. 7, 10991.
    6. Human induced pluripotent stem cells (1:300).
      Telezhkin, V. et al. (2016) Am. J. Physiol. 310, C250.
    7. Rat hippocampal neurons (1:50).
      Tang, Y. et al. (2015) Neuroscience 304, 109.
    8. Rat hippocampal neurons.
      Dupuis, J.P. et al. (2014) EMBO J. 33, 842.
    9. Rat hippocampal primary neurons (1:500).
      Mikasova, L. et al. (2012) Brain 135, 1606.
    Immunoprecipitation citations
    1. Mouse striatal tissue lysate.
      Kang, R. et al. (2019) Front. Synaptic Neurosci. 11, 3.
    Immunocytochemistry citations
    1. Rat primary hippocampal neurons.
      Schweitzer, B. et al. (2017) Sci. Rep. 7, 10991.
    2. Mouse hippocampal neurons.
      Atkin, G. et al. (2015) J. Neurosci. 35, 6165.
    3. Rat cultured hippocampal neurons (1:100).
      Swanger, S.A. et al. (2013) J. Neurosci. 33, 8898.
    4. Mouse striatal cells.
      Del Toro, D. et al. (2010) J. Neurochem. 115, 153.
    More product citations
    1. Prieto, G.A. et al. (2017) J. Neurosci. 37, 1197.
    2. Zhang, L. et al. (2017) eNeuro 4, e0175.
    3. Lin, H. et al. (2014) Neurobiol. Dis. 63, 129.
    4. Piguel, N.H. et al. (2014) Cell Rep. 9, 712.
    5. Gladding, C. et al. (2012) Hum. Mol. Genet. 21, 3739.
    6. Papouin, T. et al. (2012) Cell 150, 633.
    7. Lo, F.S. et al. (2011) Neuroscience 178, 240.
    8. Hughes, E.G. et al. (2010) J. Neurosci. 30, 5866.
    9. Nicolai, J. et al. (2010) Cell Death Dis. 1, e33.
    10. Wee, X.K. et al. (2010) Br. J. Pharmacol. 159, 449.
    11. Fontaine, R.H. et al. (2008) Proc. Natl. Acad. Sci. U.S.A. 105, 16779.
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