Overview
- Peptide (C)NTHEKRIYQSNMLNR, corresponding to amino acid residues 323-337 of rat NMDA receptor 2B (Accession Q00960). Extracellular, N-terminus.
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 NMDAR2B/GluN2B (extracellular) Blocking Peptide (#BLP-GC003).- Mouse forebrain cortex (1:200) (Fontaine, R.H. et al. (2008) Proc. Natl. Acad. Sci. U.S.A. 105, 16779.).
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.
Expression of NMDA receptor 2B in rat hippocampusImmunohistochemical 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.
Expression of NMDA receptor 2B in rat cortexImmunohistochemical 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.
- Mouse striatal cells (Del Toro, D. et al. (2010) J. Neurochem. 115, 153.).
- Rat hippocampal primary neurons (1:500) (Mikasova, L. et al. (2012) Brain 135, 1606.).
- Dingledine, R. et al. (1999) Pharmacol. Rev. 51, 7.
- Mayer, M.L. and Armstrong, N. (2004) Annu. Rev. Physiol. 66, 161.
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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.
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.
Expression 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.
Applications
Citations
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Expression of GluN2B in rat hippocampal neuronsImmunocytochemistry 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.
- Mouse striatal tissue lysate.
Kang, R. et al. (2019) Front. Synaptic Neurosci. 11, 3. - Rat primary hippocampal neuron lysate.
Schweitzer, B. et al. (2017) Sci. Rep. 7, 10991. - Mouse cortex lysate.
Lin, H. et al. (2016) Front. Cell. Neurosci. 10, 34. - Mouse brain lysate.
Wang, Y.C. et al. (2016) J. Cereb. Blood Flow Metab. 37, 980. - Mouse hippocampus lysate.
Antonelli, R. et al. (2016) J. Neurosci. 36, 5437. - Mouse brain lysate (1:2000).
Konstantoudaki, X. et al. (2016) Neuroscience 322, 333. - Mouse hippocampus lysate (1:1000).
Tindi, J.O. et al. (2015) J. Neurosci. 35, 8986. - Mouse brain lysate (1:500).
Atkin, G. et al. (2015) J. Neurosci. 35, 6165. - Mouse striatal lysate (1:500).
Dau, A. et al. (2014) Neurobiol. Dis. 62, 533.
- Rat hippocampal live, intact cells (1:100).
Afonson, P. et al. (2019) Sci. Signal. 12, eaav3577. - Mouse embryonic hippocampal neurons.
Morini, R. et al. (2018) Front. Mol. Neurosci. 11, 313. - Mouse dissociated hippocampal culture.
Altmuller, F. et al. (2017) PLoS Genet. 13, e1006684. - Rat hippocampal culture.
Ferreira, J.S. et al. (2017) eLife 6, 8986. - Rat hippocampal culture.
Joshi, P. et al. (2017) Sci. Rep. 7, 41734. - Rat primary hippocampal neurons.
Schweitzer, B. et al. (2017) Sci. Rep. 7, 10991. - Human induced pluripotent stem cells (1:300).
Telezhkin, V. et al. (2016) Am. J. Physiol. 310, C250. - Rat hippocampal neurons (1:50).
Tang, Y. et al. (2015) Neuroscience 304, 109. - Rat hippocampal neurons.
Dupuis, J.P. et al. (2014) EMBO J. 33, 842. - Rat hippocampal primary neurons (1:500).
Mikasova, L. et al. (2012) Brain 135, 1606.
- Mouse striatal tissue lysate.
Kang, R. et al. (2019) Front. Synaptic Neurosci. 11, 3.
- Rat neurons.
Sceniak, M.P. et al. (2019) J. Cell Sci. 132, jcs232892. - Rat primary hippocampal neurons.
Schweitzer, B. et al. (2017) Sci. Rep. 7, 10991. - Mouse hippocampal neurons.
Atkin, G. et al. (2015) J. Neurosci. 35, 6165. - Rat cultured hippocampal neurons (1:100).
Swanger, S.A. et al. (2013) J. Neurosci. 33, 8898. - Mouse striatal cells.
Del Toro, D. et al. (2010) J. Neurochem. 115, 153.
- Prieto, G.A. et al. (2017) J. Neurosci. 37, 1197.
- Zhang, L. et al. (2017) eNeuro 4, e0175.
- Lin, H. et al. (2014) Neurobiol. Dis. 63, 129.
- Piguel, N.H. et al. (2014) Cell Rep. 9, 712.
- Gladding, C. et al. (2012) Hum. Mol. Genet. 21, 3739.
- Papouin, T. et al. (2012) Cell 150, 633.
- Lo, F.S. et al. (2011) Neuroscience 178, 240.
- Hughes, E.G. et al. (2010) J. Neurosci. 30, 5866.
- Nicolai, J. et al. (2010) Cell Death Dis. 1, e33.
- Wee, X.K. et al. (2010) Br. J. Pharmacol. 159, 449.
- Fontaine, R.H. et al. (2008) Proc. Natl. Acad. Sci. U.S.A. 105, 16779.
