Cell Signaling Technology

Product Pathways - Neuroscience

Phospho-NMDAR2A (Tyr1246) Antibody #4206

Applications Reactivity Sensitivity MW (kDa) Source
W IP R (H) (M) Endogenous 180 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human  M=Mouse  R=Rat
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Phospho-NMDAR2A (Tyr1246) Antibody detects endogenous levels of NMDAR2A only when phosphorylated at Tyr1246. The antibody may also detect NMDAR2B when phosphorylated at the conserved Tyr1252.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr1246 of human NMDAR2A. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from rat brain, either sham-operated or 15 min ischemia, using Phospho-NMDAR2A (Tyr1246) Antibody (upper) or NMDAR2A Antibody (lower).

Background

N-methyl-D-aspartate receptor (NMDAR) forms a heterodimer of at least one NR1 and one NR2A-D subunit. Multiple receptor isoforms with distinct brain distributions and functional properties arise by selective splicing of the NR1 transcripts and differential expression of the NR2 subunits. NR1 subunits bind the co-agonist glycine and NR2 subunits bind the neurotransmitter glutamate. Activation of the NMDA receptor or opening of the ion channel allows flow of Na+ and Ca2+ ions into the cell, and K+ out of the cell (1). Each subunit has a cytoplasmic domain that can be directly modified by the protein kinase/phosphatase (2). PKC can phosphorylate the NR1 subunit (NMDAR1) of the receptor at Ser890/Ser896, and PKA can phosphorylate NR1 at Ser897 (3). The phosphorylation of NR1 by PKC decreases its affinity for calmodulin, thus preventing the inhibitory effect of calmodulin on NMDAR (4). The phosphorylation of NR1 by PKA probably counteracts the inhibitory effect of calcineurin on the receptor (5). NMDAR mediates long-term potentiation and slow postsynaptic excitation, which play central roles in learning, neurodevelopment, and neuroplasticity (6).

EphrinB2 binding to the receptor EphB leads to the activation of Src family tyrosine kinases, which phosphorylate NMDAR2B at Tyr1252, Tyr1336 and Tyr1472. In turn, phosphorylated NMDAR2B enhances the ability of the functional NMDA receptor to regulate Ca2+ influx in response to glutamate (7). The phosphorylation site of NMDAR2A at Tyr1246 is the conserved site of NMDAR2B at Tyr1252 and was independently identified at Cell Signaling Technology (CST) using PhosphoScan®, CST's MS/MS platform for phosphorylation site discovery. Phosphorylation of NMDAR2A at Tyr1246 was observed in extracts isolated from ischemic rat brain. For additional information please visit PhosphoSitePlus®, CST's modification site knowledgebase, at www.phosphosite.org.

  1. Liu, X.B. et al. (2004) J. Neurosci. 24, 8885-8895.
  2. Westphal, R.S. et al. (1999) Science 285, 93-96.
  3. Tingley, W.G. et al. (1997) J. Biol. Chem. 272, 5157-5166.
  4. Hisatsune, C. et al. (1997) J. Biol. Chem. 272, 20805-20810.
  5. Raman, I.M. et al. (1996) Neuron 16, 415-421.
  6. Makhinson, M. et al. (1999) J. Neurosci. 19, 2500-2510.
  7. Takasu, M.A. et al. (2002) Science 295, 491-495.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

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