Western blot: Detects monomeric (~95–100 kDa) and dimeric (~200 kDa) GRM3 in human, mouse, and rat brain lysates . Non-specific bands may appear with certain antibodies (e.g., N-terminal antibodies) .
Knockout validation: Antibodies show no reactivity in Grm3⁻/⁻ and Grm2⁻/⁻/Grm3⁻/⁻ mouse models, confirming specificity .
ELISA: HRP-conjugated GRM3 antibodies enable quantitative analysis with high sensitivity (e.g., detection limits in picomolar ranges) .
Immunoprecipitation: Validated for co-precipitation studies, such as GRM3 interaction with splice variants like mGlu3Δ4 .
Schizophrenia research: GRM3 antibodies identify altered receptor expression or localization in postmortem brain tissues, though no consistent changes in schizophrenia have been confirmed .
Ligand binding assays: HRP-based detection quantifies GRM3 interactions with antagonists like [³H]LY341495, revealing reduced binding in the presence of splice variants (e.g., mGlu3Δ4) .
B-cell tumors: GRM3 antibodies detect upregulated receptor expression in apoptosis-induced SP 2/0 myeloma cells treated with FK506 .
Receptor binding: HRP-conjugated antibodies validate GRM3 interactions with novel ligands (e.g., Raptin) via immunoprecipitation and saturation binding assays .
Antigen retrieval: Required for IHC in formalin-fixed tissues (e.g., sodium citrate buffer at pH 6) .
Membrane fractionation: Enrichment improves detection of GRM3 in Western blots due to its predominant membrane localization .
Blocking peptides: Pre-adsorption with immunogen peptides (e.g., TQGSHHPVTPEEC for mGlu3Δ4) confirms specificity .
Non-specific bands: Observed with some N-terminal antibodies; C-terminal antibodies show higher specificity .
Postmortem variables: GRM3 immunoreactivity declines with tissue pH and prolonged postmortem intervals .