Anti-Gephyrin Antibody

Putative glycine receptor-tubulin linker protein, MPT adenylyltransferase, MPT Mo-transferase, Gphn
    Cat #: AIP-005
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, m, r
    Immunogen
    Peptide (C)RDAIVKVKEVHDE, corresponding to amino acid residues 183-195 of rat Gephyrin (Accession Q03555). Intracellular.
    Accession (Uniprot) Number Q03555
    Gene ID 64845
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Mouse, human – 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 0.85 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 -20ºC.
    Preadsorption Control 1 µg peptide per 1 µg antibody.
    Standard quality control of each lot Western blot analysis.
    Applications: ih, wb
    May also work in: ic, ip
    Western blot
    Western blot analysis of rat brain (lanes 1 and 3) and mouse brain (lanes 2 and 4) lysates:
    1, 2. Anti-Gephyrin Antibody (#AIP-005), (1:200).
    3, 4. Anti-Gephyrin Antibody, preincubated with the control peptide antigen.
    Immunohistochemistry
    Expression of Gephyrin in rat hippocampus
    Immunohistochemical staining of rat hippocampal dentate gyrus sections using Anti-Gephyrin Antibody (#AIP-005). A. Gephyrin staining (green) appears in hilus of the dentate gyrus (arrows). B. Nuclear staining using DAPI as the counterstain (blue). C. Merge image of panel A and B.
    References
    1. Chen, J. et al. (2014) Front. Cell. Neurosci. 8, 276.
    2. Tyagarajan, S.K. and Fritschy, J.M. (2010) J. Physiol. 588, 101.
    3. Maric, H.M. et al. (2014) ACS Chem. Biol. DOI: 10.1021/cb500303a.
    4. Fritschy, J.M. et al. (2008) Trends Neurosci. 31, 257.
    5. Zacchi, P. et al. (2014) Front. Cell. Neurosci. 8, 103.
    6. Lorenzo, L.E. et al. (2014) J. Neurosci. 34, 8300.
    Scientific background

    Gephyrin is a highly conserved, intracellular scaffolding protein responsible for the post-synaptic anchoring and assembly of the inhibitory glycine and γ-aminobutyric acid type A receptors (GlyR and GABAAR); it essentially anchors the receptors, by auto-oligomerization, to the cytoskeleton1,2.

    GABAergic transmission is important for the mediation of neuronal excitability and has a crucial role in neurodevelopment, throughout the central and peripheral nervous systems (especially via GABAAR). Glycinergic neurons on the other hand control motor functions and sensory signaling as well as take part in pain transmission in the spine's dorsal horn. Importantly, gephyrin has a 1:1 stoichiometry with GlyR3,4.

    Up-to-date atomic force microscopy data of gephyrin has shown that it forms trimers with multiple conformations. It is a modular construct with three domains: an N-terminal G-, a central C- and a C-terminal E-domain. Recent work suggests also that gephyrin regulates neuronal differentiation by anchoring small cytoskeletal-related signaling molecules at unique subcellular locations2,5,6.

    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:
    Immuno-colocalization of Vesicular GABA Transporter and Gephyrin in rat spinal cord.
    Immunohistochemical staining of perfusion-fixed frozen rat spinal cord sections using Anti-Gephyrin Antibody (#AIP-005), (1:300) and Anti-Vesicular GABA Transporter-ATTO-594 Antibody (#AGT-005-AR), (1:80).  A. Gephyrin staining (green) appears in the superficial layer of the spinal dorsal horn and some cellular profiles were stained. B. Vesicular GABA Transporter staining (red) appears in superficial layer and in cell profiles. C. Merge images of A and B reveals co-localization (arrows) in superficial layers and separate expression in some cells. Cell nuclei were stained with DAPI (blue) as the counterstain.
    Last update: 04/12/2018

    Alomone Labs is pleased to offer a highly specific antibody directed against an epitope of rat Gephyrin. Anti-Gephyrin Antibody (#AIP-005) can be used in western blot and immunohistochemistry applications. It has been designed to recognize Gephyrin from rat, mouse, and human samples.

    For research purposes only, not for human use