Western Blotting (WB), Immunoprecipitation (IP), Immunohistochemistry (Frozen Sections) (IHC (fro))
Specificity
This antibody recognizes the ~180k NR2A subunit of the NMDA Receptor in Western Blots of most brain regions. No reactivity towards the NR2B and NR2C subunits. Immunolabeling is blocked by pre-adsorption of antibody with the fusion protein used to generate the antibody.
Cross-Reactivity (Details)
Species reactivity (tested):Human, Mouse and Rat.
Purification
Affinity Chromatography.
Immunogen
Fusion protein from the C-terminus of the NR2A subunit of Rat NMDA Receptor.
Western blot: 1/1000. Immunohistochemistry on Frozen Sections: 1/1000-1/2000. Immunoprecipitation: 3 μL per 200 μg lysate. Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.
Restrictions
For Research Use only
Reconstitution
Restore in 50 μL PBS (137 mM NaCl, 7.5 mM Na2HPO4, 2.7 mM KCl, 1.5 mM KH2PO4, pH 7.4) before use.
Buffer
5 mM Ammonium Bicarbonate.
Handling Advice
Avoid repeated freezing and thawing.
Storage
-20 °C
Storage Comment
Store the antibody undiluted (in aliquots) at-20 °C.
Bostrom, Majaess, Morch, White, Eadie, Christie: "Rescue of NMDAR-dependent synaptic plasticity in Fmr1 knock-out mice." in: Cerebral cortex (New York, N.Y. : 1991), Vol. 25, Issue 1, pp. 271-9, (2014) (PubMed).
Pozniak, Sengupta Ghosh, Gogineni, Hanson, Lee, Larson, Solanoy, Bustos, Li, Ngu, Jubb, Ayalon, Wu, Scearce-Levie, Zhou, Weimer, Kirkpatrick, Lewcock: "Dual leucine zipper kinase is required for excitotoxicity-induced neuronal degeneration." in: The Journal of experimental medicine, Vol. 210, Issue 12, pp. 2553-67, (2013) (PubMed).
Hicklin, Wu, Radcliffe, Freund, Goebel-Goody, Correa, Proctor, Lombroso, Browning: "Alcohol inhibition of the NMDA receptor function, long-term potentiation, and fear learning requires striatal-enriched protein tyrosine phosphatase." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 108, Issue 16, pp. 6650-5, (2011) (PubMed).
Norris, Strickland: "Modulation of NR2B-regulated contextual fear in the hippocampus by the tissue plasminogen activator system." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 104, Issue 33, pp. 13473-8, (2007) (PubMed).
Wenthold, Prybylowski, Standley, Sans, Petralia: "Trafficking of NMDA receptors." in: Annual review of pharmacology and toxicology, Vol. 43, pp. 335-58, (2003) (PubMed).
Alvestad, Grosshans, Coultrap, Nakazawa, Yamamoto, Browning: "Tyrosine dephosphorylation and ethanol inhibition of N-Methyl-D-aspartate receptor function." in: The Journal of biological chemistry, Vol. 278, Issue 13, pp. 11020-5, (2003) (PubMed).
The ion channels activated by glutamate are typically divided into two classes. Glutamate receptors that are activated by kainate and alpha-amino-3-hydroxy-5-methyl-4-isoxalone propionic acid (AMPA) are known as kainate/AMPA receptors (K/AMPAR). Those that are sensitive to N-methyl-D-aspartate (NMDA) are designated NMDA receptors (NMDAR). The NMDAR plays an essential role in memory, neuronal development and it has also been implicated in several disorders of the central nervous system including Alzheimer's, epilepsy and ischemic neuronal cell death (Grosshans et al., 2002, Wenthold et al., 2003, Carroll and Zukin, 2002). The NMDA receptor is also one of the principal molecular targets for alcohol in the CNS (Lovinger et al., 1989, Alvestad et al., 2003, Snell et al., 1996). The NMDAR is also potentiated by protein phosphorylation (Lu et al., 1999). The rat NMDAR1 (NR1) was the first subunit of the NMDAR to be cloned. The NR1 protein can form NMDA activated channels when expressed in Xenopus oocytes but the currents in such channels are much smaller than those seen in situ. Channels with more physiological characteristics are produced when the NR1 subunit is combined with one or more of the NMDAR2 (NR2 A-D) subunits.Synonyms: GRIN2A, Glutamate [NMDA] Receptor subunit epsilon-1, NMDAR2A, NR2A