GluR1 (GluA1) Overexpressed Membrane Fractions

A Positive Control for Detecting GluR1 in Western Blot Analysis
    Cat #: LX-103
  • Lyophilized Powder
  • Accession (Uniprot) Number X17184
    Origin Rat AMPA Receptor 1 (AMPA1, GRIA1, GluR1, GluA1) cDNA (X17184) was transcribed in vitro to produce full length mRNA. AMPA Receptor 1 mRNA was injected into Xenopus oocytes and protein expression was verified by detection of glutamate evoked CNQX (#C-140) or DNQX disodium salt (#D-130) sensitive ionic current. Membrane fraction was prepared from oocyte population homogenate and lyophilized. Non-injected oocytes underwent the same procedure excluding mRNA injection.
    Storage before reconstitution Lyophilized injected and non-injected membrane fractions can be stored at room temperature for 2 weeks. For longer periods, they should be stored at -20°C.
    Reconstitution Work on ice. Gently reconstitute injected and non-injected membrane fractions in 100 μl DDW (total protein concentration is 0.36 μg/μl). Centrifuge all product preparations before use (10000 x g 5 min).
    Storage after reconstitution Up to 2 weeks at -20°C. Avoid repeated thawing and freezing.
      • GluR1 (GluA1) Overexpressed Membrane Fractions
        Western blot analysis of GluR1 (GluA1) Overexpressed Membrane Fractions (#LX-103) obtained from Xenopus oocyte membrane fractions expressing rat AMPA Receptor 1 (lane 2), and non-injected oocyte lysates (lane 1):
    Note

    Lyophilized membrane fraction was reconstituted in 100 μl DDW (see below). 100 μl of Sample Buffer X2 was added (total protein concentration was 0.18 μg/μl). Boiled samples were loaded onto 10% acrylamide gel (10-40 μl/lane) and resolved using SDS-PAGE gel, followed by western blot analysis using Anti-GluR1 (GluA1) (extracellular) Antibody (#AGC-004) following product instructions.

    This product includes:
    1 x 0.1 ml lyophilized GluR1 receptor overexpressed membrane lysate
    1 x 0.1 ml lyophilized non-injected Xenopus oocyte membrane lysate

    References
      • 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.

    Last update: 14/07/2019

    GluR1 (GluA1) Overexpressed Membrane Fractions (#LX-103) are Xenopus oocyte membrane fractions overexpressing GluR1.* GluR1 (GluA1) Overexpressed Membrane Fractions are your positive control for validating Alomone Labs Anti-GluR1 (GluA1) (extracellular) Antibody (#AGC-004).

    Overexpressed Membrane Fractions are:
    ✓ Lyophilized powder
    ✓ Economical
    ✓ Shipped at room temperature (no need for dry ice and extra shipping costs)
    ✓ User-friendly & time-saving. Just add water, sample buffer and load your gel

    GluR1 (GluA1) Overexpressed Membrane Fractions, along with Anti-GluR1 (GluA1) (extracellular) Antibody, are available as part of the following kit:
    GluR1 (GluA1) Antibody and Membrane Fractions Kit (#LK-103).

    *Product includes non-injected oocyte membrane lysates

    For research purposes only, not for human use