Western blot analysis of extracts from various cell lines using Gephyrin Antibody (upper) and α/β-Tubulin Antibody #2148 (lower). As expected, gephyrin protein expression is not detected in A172 cells.
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Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 10
Gephyrin Antibody recognizes endogenous levels of total gephyrin protein.
Human, Mouse, Rat
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu517 of human gephyrin protein. Antibodies are purified by protein A and peptide affinity chromatography.
The neurotransmitters GABA and glycine activate ligand-gated chloride channels and thus mediate fast synaptic inhibition. Gephyrin is a postsynaptic, scaffolding protein anchoring type A GABA and glycine receptors to the cytoskeleton. In addition to gephyrin’s function clustering synaptic neurotransmitter receptors, it plays an essential role in the biosynthesis of the molybdenum cofactor (MoCo). Molybdenum cofactor chelates and activates sulfite oxidase, an enzyme crucial for survival (1). GSK-3β and Erk1/2 phosphorylate gephyrin at residue Ser270 and Ser268, respectively. These post-translational modifications alter the clustering of gephyrin, effecting the amplitude and frequency of GABAergic inhibitory currents (2,3). Researchers are analyzing the role of abnormal gephyrin clustering and function in major neurological, neuro-developmental and psychiatric disorders (1).
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