Anti-Gephyrin Polyclonal Antibody for WB and ELISA - Q9NQX3

Core Product Specifications and Parameters

Biological Background: Gephyrin Function and Localization

• Gephyrin is a multifunctional protein primarily known as a microtubule-associated protein that anchors inhibitory neurotransmitter receptors to the cytoskeleton at postsynaptic sites.
• It serves as a major instructive molecule at inhibitory synapses, clustering both glycine receptors (GLYR) and GABA type A receptors, thereby stabilizing synaptic architecture. Related references: PMID:25025157, PMID:26613940
• Beyond its structural role, gephyrin catalyzes two steps in the biosynthesis of the molybdenum cofactor (MoCo), involving adenylation of molybdopterin followed by molybdate insertion and AMP release.
• The GPHN gene encodes several isoforms through alternative splicing, contributing to functional diversity in both neuronal and non-neuronal tissues.
• Subcellularly, gephyrin localizes to the postsynaptic cell membrane, dendrites, and postsynaptic density in neurons, but is also found in the cytoplasm, cytosol, and associated with the cytoskeleton in other cell types.
• Post-translational modifications include palmitoylation and phosphorylation, which regulate its membrane association and clustering activity.
• Disease-associated mutations in GPHN have been linked to neurological disorders, emphasizing its critical role in synaptic function.

Experimental Guidance and Technical Tips

• The immunogen spans the C-terminal region (aa 520–769) of human gephyrin, which encompasses sequences involved in receptor clustering; therefore, this antibody may detect isoforms containing this domain.
• For Western blot, a band around 80 kDa is expected; however, alternative splicing and post-translational modifications might result in additional bands. Validate the specificity using appropriate controls (e.g., knockdown or knockout lysates).
• The antibody cross-reacts with mouse and rat gephyrin; consider sequence alignment of the immunogen region to confirm homology for other species before use.
• This polyclonal antibody is suitable for ELISA; standard optimization of coating antigen concentration and detection conditions is recommended.
• For immunohistochemical applications (not validated in the product's applications), antigen retrieval methods may be necessary due to the tight synaptic localization; perform pilot experiments.

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