Detection of GRIN2C/NMDAR2C by Western Blot.
Western blot of 10 μg of rat brain (cerebellum) lysate showing specific immunolabeling of the ~140 kDa NR2C subunit of the NMDA Receptor. The labeling by the antibody was blocked by the N-Terminal NR2C subunit NMDA Receptor fusion protein used as the antigen.
Preparation and Storage
This antibody should be reconstituted in 50 μL phosphate buffered saline (137 mM NaCl, 7.5 mM Na2HPO4, 2.7 mM KCl, 1.5 mM KH2PO4, pH 7.4) before use.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
The lyophilized product is stable at ≤ -20° C for at least 1 year. After reconstitution the antibody should be aliquoted and stored at ≤ -20° C.
NMDA (N-Methyl D-Aspartate) receptors are members of the glutamate receptor family of ligand-gated ion channels. The functional NMDA receptor (NMDAR) is a 650 - 850 kDa heteromultimer of at least two NR2 (NMDAR2) subunits and two NR1 subunits. NR2 subunits determine overall NMDAR characteristics such as conductance and Mg++ sensitivity. In addition, NR2 subunits mediate NMDAR clustering and synaptic localization through cytoplasmic interaction with PSD-95/SAP90 family members. Upon glutamate binding to NR2, and glycine binding to NR1, the NMDA channel is opened, allowing calcium and sodium influx into the cell. There are four genes that code for NR2 subunits (NR2A-D). The two (or three) NR2 subunits making up the NMDAR may be homodimers or heterodimers. The 2C form generates a low conductance NMDAR. This receptor is particularly abundant in the cerebellum and thalamus. Rat NR2C is a 180 kDa, 1237 amino acid (aa), three transmembrane (TM) glycoprotein that contains a 535 aa extracellular domain (ECD) and a 401 aa cytoplasmic region. The molecule is described as 4-TM. However, the second-TM segment is only partial (or reenterant), and this makes the C-terminus intracellular. The loop connecting TM segments 3 and 4, plus the N-terminal ECD likely constitute the glutamate binding site. Rat NR2C shares 97% and 88% aa sequence identity to mouse and human NR2C, respectively.
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Prybylowski, K. and R.J. Wenthold (2004) J. Biol. Chem. 279:9673.
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