Polyclonal Antibody to NMDA NR1, Splice Variant C2

Background
The ion channels activated by glutamate are typically divided into two classes. Those that are sensitive to N-methyl-D-aspartate (NMDA) are designated NMDA receptors (NMDAR). NMDARs play an essential role in memory, neuronal development and have also been implicated in several disorders of the central nervous system including Alzheimer’s disease, epilepsy and ischemic neuronal cell death. Increased membrane surface expression of the NR1 subunit of the receptor has been associated with synaptic plasticity. There are a number of different splice variants of the NR1 subunit. Differential splicing of three exons in the NR1 subunit generates up to eight NR1 splice variants and 7 of these have been identified in cDNA libraries. These exons encode a 21 amino acid N-terminal domain (N1) and adjacent sequences in the C-terminus (C1 and C2). Splicing out the C2 cassette eliminates the first stop codon and produces a new reading frame that generates a new sequence of 22 amino acids (C2'). Considerable attention has been focused on the distribution and expression of these splice variants as they may affect the functional properties and regulation of the NMDAR.

Antigen
A synthetic peptide from derived from the C2' splice variant insert of NMDAR NR1 was used as immunogen. The antibody was affinity purified on a column made with the peptide used as the antigen.

Application Notes
Specific for the NR1-subunit of the NMDA Receptor containing the C2’ Splice Variant Insert in Western blots of rat brain extracts. Does not recognize the NR1-subunits of the NMDA Receptor that do not contain the C2’ Insert. Also recognizes the NR-1-subunit, C2’ Splice Variant Insert in mouse brain extracts. Antibody dilutions and tissue load should be based on tissue type and expected expression level.

Reference
1. Grosshans DR, Clayton DA, Coultrap SJ, Browning MD. 2002. LTP leads to rapid surface expression on NMDA not AMPA receptors in adult rat CA1.  Nature Neurosci. 5:27 - 33.
2. Luo JH, Wang YH, Yasuda RP,  Dunah AW, Wolfe BB. 1997. The majority of N-methyl-D-aspartate receptor complexes in adult rat cerebral cortex contain at least three different subunits (NR1/NR2A/NR2B). Mol. Pharmacol. 51:79 - 86.
3. Zukin RS and Bennett MVL. 1995. Alternatively spliced isoforms of the NMDARI receptor subunit. Trends Neurosci., 18:306 - 313.
4. Laurie DJ. and Seeburg PH.  1994. Regional and developmental heterogeneity in splicing of the rat brain NMDAR1 mRNA. J. Neurosci. 14:3180 - 3194.
5. Foldes RL, Rampersad V, Kamboj RK. 1994. Cloning and sequence analysis of additional splice variants encoding human N-methyl-D-aspartate receptor (hNR1) subunits. Gene 147:303 - 304.