Guinea pig Anti-GluR1 (GluA1) (extracellular) Antibody

AMPA Receptor 1, Glutamate receptor 1, Ionotropic glutamate receptor 1, AMPA-selective glutamate receptor 1, GRIA1, GluR-A, GluR-K1
    Cat #: AGP-009
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Guinea pig
    Reactivity: h, m, r
    Immunogen
    Peptide RTSDSRDHTRVDWKR(C), corresponding to amino acid residues 271-285 of rat GluR1 (Accession P19490). Extracellular, N-terminus.
    Accession (Uniprot) Number P19490
    Gene ID 50592
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Mouse - identical; human, pig, dog - 13/14 amino acid residues identical.
    Purity Affinity purified on immobilized antigen.
    Formulation Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN3.
    Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
    Reconstitution 25 µl, 50 µl or 0.2 ml double distilled water (DDW), depending on the sample size.
    Antibody concentration after reconstitution 0.8 mg/ml.
    Storage after reconstitution The reconstituted solution can be stored at 4°C 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 x 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.
    Applications: ic, ih, lci, wb
    May also work in: ifc, ip
    Western blot
    Western blot analysis of rat (lanes 1 and 3) and mouse (lanes 2 and 4) brain lysates:
    1,2. Guinea pig Anti-GluR1 (GluA1) (extracellular) Antibody (#AGP-009), (1:200).
    3,4. Guinea pig Anti-GluR1 (GluA1) (extracellular) Antibody, preincubated with the control peptide antigen.
    Immunohistochemistry
    Expression of GluR1 in rat hippocampus
    Immunohistochemical staining of perfusion-fixed frozen rat brain sections using Guinea pig Anti-GluR1 (GluA1) (extracellular) Antibody (#AGP-009), (1:400), followed by anti-rabbit-Cy2 antibody (green). GluR1 staining appears in neuronal outlines (horizontal arrows) and in the inner molecular layer of the dentate gyrus (vertical arrow). Nuclei are stained with DAPI (blue).
    Live cell imaging / Immunocytochemistry
    Immuno-colocalization of GluR1 and Vesicular GABA Transporter in human U-87 MG cells
    Immunocytochemical staining of human glioblastoma U-87 MG. Extracellular staining of live intact cells with Guinea pig Anti-GluR1 (GluA1) (extracellular) Antibody (#AGP-009), (1:25), followed by goat anti-guinea pig-AlexaFluor-488 secondary antibody (green). Cells were subsequently fixed, permeabilized and labeled with Anti-Vesicular GABA Transporter Antibody (#AGT-005), (1:200), followed by goat anti-rabbit-AlexaFluor-594 secondary antibody (red). Representative merged images of the double labeled cells are shown in A and B.
    References
    1. Dingledine, R. et al. (1999) Pharmacol. Rev51, 7.
    2. Sheng, M. et al. (2001) Cell. 105, 825.
    3. Song, I. et al. (2002) Trends. Neurosci25, 578.
    Scientific background

    AMPA receptors are members of the glutamate receptor family of ion channels that also include the NMDA and Kainate receptors. The three subfamilies are named after the original synthetic agonists that were identified as selective ligands of each family.

    The α-amino-3-hydroxy-5-methyl-4-isoazolepropionic acid (AMPA) receptor subfamily includes four members AMPA1-AMPA4 that are also known as GluR1-GluR4 respectively.

    The functional AMPA channel is believed to be a tetramer, with most neuronal AMPA receptors being actually heterotetramers composed of AMPA1 plus AMPA2 or AMPA2 plus AMPA3, although homotetramers can also be found.

    AMPA receptors are permeable to cations Na+, K+ and Ca2+. The Ca2+ permeability is dependent on the presence of AMPA2: whenever this subunit is present, the channel will be impermeable to Ca2+. The Ca2+ permeability of the AMPA2 subunit is determined by the presence of an arginine (R) at a critical site in the pore loop instead of a glutamine (Q) present in the same site in the other AMPA subunits. A post-transcriptional process known as RNA editing determines the presence of this R. Since most AMPA2 subunits in the adult brain have undergone RNA editing and most AMPA receptors contain the AMPA2 subunit, most native AMPA receptors will be impermeable to Ca2+.

    Gating of AMPA receptors by glutamate is extremely fast and therefore the AMPA receptors mediate most excitatory (depolarizing) currents in the brain during basal neuronal activity. The depolarization caused by the activation of post-synaptic AMPA receptors is necessary for the activation of NMDA receptors that will open only in the presence of both glutamate and a depolarized membrane.

    Synaptic strength, defined as the level of post-synaptic depolarization, can be long term (hence the term long term potentiation, LTP) and therefore induce changes in signaling and protein synthesis in the activated neuron. These changes are associated with memory formation and learning.

    Changes in synaptic strength are thought to involve rapid movement of the AMPA receptors in and out of the synapses and a great deal of effort has focused in understanding the mechanisms that govern AMPA receptor trafficking.

    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
    Image & Title: Guinea pig Anti-GluR1 (GluA1) (extracellular) Antibody
    Expression of GluR1 in mouse olfactory bulb.Immunohistochemical staining of mouse olfactory bulb sections using Guinea pig Anti-GluR1 (GluA1) (extracellular) Antibody (#AGP-009). GluR1 staining (green) is detected in the glomerular and external plexiform layers (EPL), (right panel). GluR1 co-localizes with GFAP in periglomerular astrocytes and their processes in the neuropil (merged panel).Adapted from Droste, D. et al. (2017) Sci. Rep. 7, 44817. with permission of Nature Publishing Group.
    Last update: 29/11/2018

    Alomone Labs is pleased to offer an antibody against an extracellular epitope the rat ionotropic glutamate receptor 1. Guinea pig Anti-GluR1 (GluA1) (extracellular) Antibody (#AGP-009) raised in guinea pig can be used in western blot, immunohistochemistry and immunocytochemistry applications. It has been designed to recognize GluR1 from human, mouse and rat samples. The antigen used to immunize guinea pigs is the same as Anti-GluR1 (GluA1) (extracellular) Antibody (#AGC-004) raised in rabbit. Our line of guinea pig antibodies enables more flexibility with our products such as immuno-colocalization studies, immunoprecipitation, etc.

    For research purposes only, not for human use
    Citations
    Immunohistochemistry citations
    1. Mouse cerebellum and olfactory bulb sections (1:200).
      Droste, D. et al. (2017) Sci. Rep. 7, 44817.
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