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Anti-Glycine Receptor α4 (extracellular) Antibody

Glycine receptor subunit alpha-4, GLRA4
Cat #: AGR-015
Alternative Name Glycine receptor subunit alpha-4, GLRA4
Lyophilized Powder yes
Type: Polyclonal
Host: Rabbit
Reactivity: h, m, r
Immunogen
  • Peptide (C)KEDVKSGLKGSQPMS, corresponding to amino acid residues 28-42 of mouse GlyRα4 (Accession Q61603). Extracellular, N-terminus.
Accession (Uniprot) Number Q61603
Gene ID 14657
Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
Homology Rat - 14/15 amino acid residues identical; human - 13/15 amino acid residues identical.
RRID AB_2340974.
Purity Affinity purified on immobilized antigen.
Form Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN3.
Isotype Rabbit IgG.
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 0.58 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).
Standard quality control of each lot Western blot analysis.
Applications: ic, if, lci, wb
May also work in: ifc*, ih*, ip*
Western blot
  • Western blot analysis of rat eye (lanes 1 and 3) and ARPE-19 (lanes 2 and 4) lysates:
    Western blot analysis of rat eye (lanes 1 and 3) and ARPE-19 (lanes 2 and 4) lysates:
    1,2. Anti-Glycine Receptor α4 (extracellular) Antibody (#AGR-015), (1:500).
    3,4. Anti-Glycine Receptor α4 (extracellular) Antibody, preincubated with Glycine Receptor α4 (extracellular) Blocking Peptide (#BLP-GR015).
Live cell imaging / Immunocytochemistry
  • Expression of GlyRα4 in human retinal epithelium (ARPE-19)
    Expression of GlyRα4 in human retinal epithelium (ARPE-19)
    Cell surface detection of GlyRα4 in intact living human ARPE-19 cells. A. Extracellular staining of cells using Anti-Glycine Receptor α4 (extracellular) Antibody (#AGR-015), (1:50) followed by goat anti-rabbit-AlexaFluor-594 secondary antibody. B. Merge image of A and live view of the cells.
References
  1. Meyer, G. et al. (1995) Neuron 15, 563.
  2. Lynch, J.W. et al. (2009) Neuropharmacology 56, 303.
  3. Bode, A. and Lynch, J.W. (2014) Mol. Brain 7, 2.
  4. Harvey, R.J. et al. (2000) Eur. J. Neurosci. 12, 994.
  5. Schaefer, N. et al. (2012) Front. Mol. Neurosci. 5, 98.
Scientific background

The amino acid glycine mediates neuronal inhibition by activating glycine receptors (GlyRs), which are ligand-gated chloride channels of the nicotinic acetylcholine receptor superfamily.  GlyRs are known to mediate postsynaptic inhibition in spinal cord, brain stem and some higher brain regions1. GlyR is a member of the pentameric Cys-loop ion channel receptor family. Functional Cys-loop receptors comprise homomeric or heteromeric pentameric oligomers with each of the five subunits arranged symmetrically in a ring around a central ion-conducting pore. Several developmentally and regionally regulated GlyR isoforms exist, which result from a differential expression of the GlyR α (α1- α4) and β subunit genes. Each GlyR subunit comprises a large extracellular amino-terminal domain that harbors the ligand-binding sites and the eponymous Cys-loop. This connects to a bundle of four α-helical transmembrane domains (labelled M1–M4) with a large intracellular domain between M3 and M4 and a short extracellular C-terminal tail2. The human α4 subunit is considered a pseudogene on the grounds that it incorporates a premature stop codon upstream of the final TM4 domain3.

The α4 subunit has been found in the adult mouse retina where it clusters together with synaptic markers such as bassoon or gephyrin. Furthermore, GlyR α4 transcripts have been detected in the white matter tract of the developing spinal cord from mice. The avian α4 mRNA was also found in dorsal root ganglia and genital ridge 4.

Although the function of GlyR α4 at its sites of expression is still unknown, homomeric α4 receptors are able to form fully functional glycine-gated Cl channels in vitro that can be antagonized with strychnine5.

Application key:

CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, IF- Immunofluorescence, 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: 23/08/2020

Alomone Labs is pleased to offer a highly specific antibody directed against an extracellular epitope of the mouse glycine receptor α4. Anti-Glycine Receptor α4 (extracellular) Antibody (#AGR-015) can be used in western blot and live cell imagine applications. The antibody recognizes an extracellular epitope and thus is ideal for detecting GlyRα4 in living cells. It has been designed to recognize GlyRα4 from human, rat and mouse samples.

For research purposes only, not for human use
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