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Affinity purified polyclonal antibody
- SPECIFICATION
- CITATIONS
- PROTOCOLS
- BACKGROUND
Application 
| WB |
|---|---|
| Primary Accession | P19493 |
| Reactivity | Human, Mouse, Rat |
| Host | Rabbit |
| Clonality | Polyclonal |
| Calculated MW | 100758 Da |
| Homology | Mouse, human - identical. |
| Gene ID | 29629 |
|---|---|
| Other Names | Glutamate receptor 4, GluR-4, GluR4, AMPA-selective glutamate receptor 4, GluR-D, Glutamate receptor ionotropic, AMPA 4, GluA4, Gria4, Glur4 |
| Related products for control experiments | Control peptide antigen (supplied with the antibody free of charge). |
| Target/Specificity | Peptide (C)KLMDRWKKLDQRE, corresponding to amino acid residues 274-286 of rat AMPA Receptor 4 (Accession P19493). Extracellular, N-terminus. |
| Dilution | WB~~1:200-1:2000 IHC~~1:100 |
| Peptide Confirmation | Confirmed by amino analysis. |
| Format | Affinity purified antibody, lyophilized powder |
| Reconstitution | 50 µl or 0.2 ml deionozed water, depending on the sample size. |
| Antibody Concentration After Reconstitution | 0.6 mg/ml. |
| Buffer After Reconstitution | Phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.025% NaN3. |
| Storage Before Reconstitution | Lyophilized powder can be stored intact at room temperature for several weeks. For longer periods, it should be stored at -20°C. |
| Storage After Reconstitution | The reconstituted solution can be stored at 4ºC for up to 2 weeks. For longer periods, small aliquots should be stored at -20ºC or below. Avoid multiple freezing and thawing. The further dilutions should be made using a carrier protein such as BSA (1%). Centrifuge all antibody preparations before use (10000 × g 5 min). |
| Control Antigen Storage Before Reconstitution | Lyophilized powder can be stored intact at room temperature for several weeks. For longer periods, it should be stored at -20°C. |
| Control Antigen Reconstitution | 100 µl water. |
| Control Antigen Storage After Reconstitution | -20ºC. |
| Preadsorption Control | 1 µg peptide per 1 µg antibody. |
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Provided below are standard protocols that you may find useful for product applications.
Background
Glutamate is the major excitatory neurotransmitter and modulates its effects via ionotropic and metabotropic receptors which are different in their molecular, biochemical, pharmacological and physiological properties1. The ionotropic ligand-gated ion channel glutamate receptors are classified into three major subtypes, AMPA, kainate and NMDA receptors according to their selective agonist. AMPA receptors consist of four closely related genes with about 70% homology that encode the four subunits GluR1-4. Their expression is known to be developmentally regulated2 and are also known to undergo alternative splicing. In fact, all four receptors have two splice variants, termed flip and flop, in an extracellular region adjacent to the M4 transmembrane region3. These splice variants are also developmentally regulated such that flip is expressed before birth and remains highly expressed throughout adulthood whereas flop expression is postnatal and reaches flip’s level in adulthood4. In addition to their expression regulation, flip and flop also differ in their activation/inactivation kinetics. Each AMPA receptor contains 3 membrane spanning domains (M1, M3 and M4), while the fourth hydrophobic domain (M2) is a reentering cytoplasmic loop that forms part of the channels pore1. Different stoichiometries of these receptors form tetrameric structures to eventually form functional receptors5. AMPA receptors are highly expressed in the brain. However, the GluR4 subunit is present in lower amounts in the CNS, except in the reticular thalamic neuclei and the cerebellum1. The primary depolarization in glutamate neurotransmission is mediated by AMPA receptors, which are also key players in synaptic plasticity, learning and memory formation. The cellular distribution and post-translational modifications like phosphorylation are important for the activity of AMPA receptors and are thought to play a role in the long lasting and activity-dependent changes in synaptic strength6-9. Abgent is pleased to offer a highly specific antibody directed against an epitope located in the extracellular N-terminal domain of the rat AMPA Receptor 4. Anti-AMPA Receptor 4 (GluA4) (extracellular) antibody (#AG1253) can be used in western blot and immunohistochemical applications, and has been designed to recognize AMPA receptor 4 from mouse and human samples.
References
References 1. Santos, S.D. et al. (2009) Neuroscience 158, 105. 2. Palmer, C.L. et al. (2005) Pharmacol. Rev. 57, 253. 3. Monyer, H. et al. (1991) Neuron 6, 779. 4. Dingledine, R. et al. (1999) Pharmacol. Rev. 51, 7. 5. Malinow, R et al. (2000) Curr. Opin. Neurobiol. 10, 352. 6. Rumpel, S. et al. (2005) Science 308, 83. 7. Morris, R.G. (2006) Eur. J. Neurosci. 23, 2829. 8. Pastalkova, E. et al. (2001) Science 313, 1141. 9. Whitlock, J.R. et al. (2006) Science 313, 1093.
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