Anti-EAAT2 (GLT-1) (extracellular)-ATTO-633 Antibody

Excitatory amino acid transporter 2, Sodium-dependent glutamate/aspartate transporter 2, SLC1A2
Cat #: AGC-022-FR
Alternative Name Excitatory amino acid transporter 2, Sodium-dependent glutamate/aspartate transporter 2, SLC1A2
  • Lyophilized Powder
  • Antigen Incl.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, m, r
      • Peptide (C)KQLGPGKKNDEVS, corresponding to amino acid residues 151-163 of rat EAAT2 (Accession P31596). 2nd extracellular loop.
    Accession (Uniprot) Number P31596
    Gene ID 29482
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Mouse, human - identical.
    Purity Affinity purified on immobilized antigen.
    Form Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN3.
    Label ATTO-633. Maximum absorption 629 nm; maximum fluorescence 657 nm. The fluorescence is excited most efficiently in the 610 - 645 nm range. This label is analogous to the dyes Alexa 647, Alexa 633 and Cy5 and can be used for direct flow cytometry (FACS) using the He:Ne laser.
    Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
    Reconstitution 50 µl double distilled water (DDW).
    Antibody concentration after reconstitution 1 mg/ml.
    Storage after reconstitution The reconstituted solution can be stored at 4°C, protected from the light, for up to 1 week. For longer periods, small aliquots should be stored at -20°C. Avoid multiple freezing and thawing. 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 2 weeks. For longer periods, it should be stored at -20°C.
    Control antigen reconstitution 100 µl double distilled water (DDW).
    Control antigen storage after reconstitution -20°C.
    Preadsorption Control 1 µg peptide per 1 µg antibody.
    Standard quality control of each lot Western blot analysis (unlabeled antibody, #AGC-022), and immunohistochemistry (labeled antibody).
    Applications: ih
    May also work in: ic, lci
      • Expression of EAAT2 in rat hippocampus
        Immunohistochemical staining of perfusion-fixed frozen rat hippocampus sections using Anti-EAAT2 (GLT-1) (extracellular)-ATTO-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.
      • 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.

    Application key:

    CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, IFC- Indirect flow cytometry, IHC- Immunohistochemistry, IP- Immunoprecipitation, LCI- Live cell imaging, N- Neutralization, WB- Western blot

    Species reactivity key:

    H- Human, M- Mouse, R- Rat
    Last update: 12/04/2019

    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-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-633 Antibody has been tested in immunohistochemical staining and is specially suited for experiments requiring simultaneous labeling of different markers.

    For research purposes only, not for human use