Anti-NMDAR2B (GluN2B) (extracellular)-ATTO-594 Antibody

NMDA receptor 2B, Ionotropic glutamate receptor subunit ε2, N-methyl-D-aspartate receptor subunit 2B, GRIN2B, NR2B
    Cat #: AGC-003-AR
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, m, r
    Immunogen
    Peptide (C)NTHEKRIYQSNMLNR, corresponding to amino acid residues 323-337 of rat NMDAR2B (Accession Q00960). Extracellular, N-terminus.
    Accession (Uniprot) Number Q00960
    Gene ID 24410
    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, 0.025% NaN3.
    Label ATTO-594. Maximum absorption 601 nm; maximum fluorescence 627 nm. The fluorescence is excited most efficiently in the 580 - 615 nm range. This label is related to the Rhodamine dyes and can be used with filters used to detect Texas Red and Alexa-594.
    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-003), and immunohistochemistry (labeled antibody).
    Applications: ih
    May also work in: ic, lci
    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 NMDAR2B (NR2B) in rat DRG
    Immunohistochemical staining of rat dorsal root ganglia (DRG) frozen sections using Anti-NMDAR2B (GluN2B) (extracellular)-ATTO-594 Antibody (#AGC-003-AR), (red), (1:50). Staining is present in neuronal cell bodies. Hoechst 33342 is used as the counterstain (blue).
    References
    1. Dingledine, R. et al. (1999) Pharmacol. Rev51, 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 diseases.

    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-NMDAR2B (GluN2B) (extracellular)-ATTO-594 Antibody
    Immuno-colocalization of PSD-95 and GluN2B in rat brain
    Immunohistochemical staining of perfusion-fixed frozen rat parietal cortex sections using Anti-PSD-95 Antibody (#APZ-009), (1:400) and Anti-NMDAR2B (GluN2B) (extracellular)-ATTO-594 Antibody (#AGC-003-AR), (1:60). A. PSD-95 staining (green). B. The same sections were stained for GluN2B (red). C. Merge of the two images shows several cells expressing both proteins (arrows point to several examples). Nuclei are stained with DAPI (blue).

    Last update: 24/10/2018

    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.

    Anti-NMDAR2B (GluN2B) (extracellular)-ATTO-594 Antibody (#AGC-003-AR) is directly labeled with an ATTO-594 fluorescent dye. ATTO dyes are characterized by strong absorption (high extinction coefficient), high fluorescence quantum yield, and high photo-stability. The ATTO-594 fluorescent label belongs to the class of Rhodamine dyes and can be used with fluorescent equipment typically optimized to detect Texas Red and Alexa-594. Anti-NMDAR2B (GluN2B) (extracellular)-ATTO-594 Antibody has been tested in immunohistochemical applications and is specially suited to experiments requiring simultaneous labeling of different markers.

    For research purposes only, not for human use