Overview
- Peptide (C)KQLGPGKKNDEVS, corresponding to amino acid residues 151-163 of rat EAAT2 (Accession P31596). 2nd extracellular loop.
Expression of EAAT2 in rat hippocampusImmunohistochemical staining of perfusion-fixed frozen rat hippocampus sections using Anti-EAAT2 (GLT-1) (extracellular)-ATTO Fluor-633 Antibody (#AGC-022-FR), (1:50). A. EAAT2 (magenta) is detected in clusters (arrows) along the sub-granular layer of the dentate gyrus. B. DAPI counterstain is used to visualize cell nuclei (blue). C. Merge of A and B. G = granule layer.
Cell surface detection of EAAT2 (GLT-1) by direct flow cytometry in live intact Rat pheochromocytoma PC12 cells:___ Cells.
___ Cells + Rabbit IgG Isotype Control-ATTO-633.
___ Cells + Anti-EAAT2 (GLT-1) (extracellular)-ATTO Fluor-633 Antibody (#AGC-022-FR), (5µg).
- Tzingounis, A.V. and Wadiche, J.I. (2007) Nat. Rev. Neurosci. 8, 935.
- Beart, P.M. and O’Shea, R.D. (2007) Br. J. Pharmacol. 150, 5.
L-Glutamate (Glu) is an abundant amino acid that functions as the major excitatory neurotransmitter in the central nervous system. However, excess of Glu in the extracellular synaptic milieu leads to neuronal cell death by a process known as excitotoxicity.
The extracellular levels of Glu are regulated by a family of high affinity plasma membrane transporters called excitatory amino acid transporters (EAATs) which are responsible for the re-uptake of Glu into the cells1,2.
The EAAT family includes five members (EAAT1-EAAT5) that are members of the solute carrier family 1 (SLC1) of Na+-dependent transporters that also includes the neutral amino acid transporters ASCT1 and ASCT2.
The Glu transporters present an unusual topology of eight transmembrane domains with two re-entrant loops and intracellular N- and C- termini. The transporter is likely assembled as a trimer where each monomer is a functional unit capable of binding the Glu substrate.
The transport of Glu into the cells by the EAAT transporters is coupled to the Na+ and K+ electrochemical gradient as a driving force. Hence, the uptake of Glu is dependent on the co-transport of three Na+ and one H+ ions, and the counter transport of one K+ ion.
In addition, to the well documented Glu uptake, the EAAT transporters show a Glu-independent Cl- conductance. The physiological significance of the Cl- current through the EAATs is currently unknown1,2.
EAAT2 (also known as Glutamate Transporter-1, GLT-1) as well as EAAT1, is expressed predominantly in glia cells, while EAAT3, EAAT4 and EAAT5 are mostly expressed in neurons.
As mentioned earlier, EAAT transporters represent the only (significant) mechanism for removal of glutamate from the extracellular fluid and hence are essential for the long-term maintenance of low and non-toxic concentrations of glutamate and the preservation of normal excitatory synaptic transmission.
In addition to Glu uptake, the glutamate transporters provide glutamate for the synthesis of g-Aminobutyric acid (GABA), glutathione and protein, suggesting an interactive role between EAATs and cellular metabolism1,2.
Dysregulation of EAAT activities has been implicated in several neurodegenerative disorders such as Alzheimer’s disease, traumatic brain injury, epilepsy and schizophrenia, suggesting that EAATs can be a useful target for the treatment of these conditions1,2.
Anti-EAAT2 (GLT-1) (extracellular) Antibody (#AGC-022) is a highly specific antibody directed against an epitope of the rat Excitatory amino acid transporter 2. The antibody can be used in western blot, live cell imaging, and immunohistochemistry applications. It has been designed to recognize EAAT2 from human, rat, and mouse samples.
Anti-EAAT2 (GLT-1) (extracellular)-ATTO Fluor-633 Antibody (#AGC-022-FR) is directly labeled with an ATTO-633 fluorescent dye. ATTO dyes are characterized by strong absorption (high extinction coefficient), high fluorescence quantum yield, and high photo-stability. ATTO 633 has a maximum absorption at 629 nm and a maximum fluorescence at 657 nm. The fluorescence is excited most efficiently in the range 610 to 645 nm. This label is analogous to the well-known dyes Alexa 647, Alexa 633 and Cy5. Anti-EAAT2 (GLT-1) (extracellular)-ATTO Fluor-633 Antibody has been tested in immunohistochemical staining and is specially suited for experiments requiring simultaneous labeling of different markers.
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