|Calculated MW||56772 Da|
|Homology||Mouse, human - 13/14 amino acid residues identical.|
|Other Names||Excitatory amino acid transporter 3, Excitatory amino-acid carrier 1, Sodium-dependent glutamate/aspartate transporter 3, Solute carrier family 1 member 1, Slc1a1, Eaac1, Eaat3|
|Related products for control experiments||Control peptide antigen (supplied with the antibody free of charge).|
|Target/Specificity||Peptide (C)EPTILDNEDSDTKK, corresponding to amino acid residuesֲ 487-500 of rat EAAT3 (Accession P51907). Intracellular, C-terminus.|
|Peptide Confirmation||Confirmed by mass-spectrography and amino acid analysis.|
|Format||Affinity purified antibody, lyophilized powder|
|Reconstitution||50 µl or 0.2 ml deionized water, depending on the sample size.|
|Antibody Concentration After Reconstitution||0.8 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.
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 as well as EAAT1 are expressed predominantly in glia cells, while EAAT3 (Excitatory Amino Acid Transporter 3), EAAT4 and EAAT5 are mostly expressed in neurons. As mentioned, 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. Abgent is pleased to offer a highly specific antibody directed against an epitope located in the intracellular C-terminal domain of the rat EAAT3 Transporter. Anti-Excitatory Amino Acid Transporter 3 antibody (#AG1250) can be used in Western blot and immunohistochemical applications, and recognizes EAAT3 from rat and mouse samples.
References 1. Tzingounis, A.V. and Wadiche, J.I. (2007) Nat. Rev. Neurosci. 8, 935. 2. Beart, P.M. and O’Shea, R.D. (2007) Br. J. Pharmacol. 150, 5.
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