GluR1 (phospho Ser831) Antibody

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) while those activated by α -amino-3-hydroxy-5-methyl-4-isoxalone propionic acid (AMPA) are known as AMPA receptors (AMPAR) . The AMPAR are comprised of four distinct subunits GluR1-4 and they play key roles in virtually all excitatory neurotransmission in the brain. The GluR1 subunit is widely expressed throughout the nervous system. GluR1 is also potentiated by phosphorylation at Ser831. In addition, phosphorylation of this site has been linked to synaptic plasticity as well and learning and memory.

GluR1 (Ser831) polyclonal antibody was raised against a synthetic phosphopeptide corresponding to amino acids residues surrounding the phospho-Ser831 of rat GluR1.

For long term storage –80˚ C is recommended, but shorter term storage at –20˚ C is also acceptable as aliquots may be taken without freeze/thawing due to the presence of 50% glycerol. Stable for one year.

Immunolabeling of the GluR1 protein in Western blots of rat brain is blocked by the Ser831 phosphopeptide used as antigen but not by the corresponding non-phosphopeptide. The antibody is purified by sequential chromatography on phospho- and non-phosphopeptide affinity columns. Antibody dilutions and tissue load should be based on tissue type and expected phosphorylation state. Initial recommended range of dilutions: 1:500 to 1:2000. Applications include Dot Blots (DB) and Western Blot (WB) . Immunohistochemistry (IHC) has not yet been determined. Human, mouse and rat have 100% amino acid sequence identity with the antigen used to raise the antibody. When internally tested under ideal conditions the working dilutions were 1:1000 for DB and WB.

Lee, H.K., Takamiya, K., Han, J.S., Man, H.Y., Kim, C.H., Rumbaugh, G., Yu, S., Ding, L., He, C., Petralia, R.S., Wenthold, R.J., Gallagher, M. and Huganir, R.L., "Phosphorylation of the AMPA receptor GluR1 subunit is required for synaptic plasticity and retention of spatial memory, " Cell 112 (2003) 631 - 643.

Banke, T.G., Bowie, D., Lee, H.K., Huganir, R.L., Schousboe, A., and Traynelis, S.F. "Control of GluR1 AMPA receptor function by cAMP-dependent protein kinase, " J. Neurosci. 20 (2000) 89 - 102.

Barria, A., Muller, D., Derkach, V., Griffith, L.C. and Soderling, T.R., "Regulatory phosphorylation of AMPA-type glutamate receptors by CaM-KII during long-term potentiation, " Science 276 (1997) 2042 - 2045.

Roche, K.W., O'Brien, R.J., Mammen, A.L., Bernhardt, J., and Huganir, R.L., "Characterization of multiple phosphorylation sites on the AMPA receptor GluR1 subunit, " Neuron, 16 (1996) 1179 - 1188.