Anti-Metabotropic Glutamate Receptor Type 1 (mGluR1) Antibody

The metabotropic glutamate receptors (mGluRs) represent a gene family of G-protein-coupled receptors, all of which bind the amino acid glutamate (the major excitatory neurotransmitter of the nervous system) but differ in their primary amino acid structure, their pharmacology, and their distribution in the peripheral and central nervous systems. In general, mGluR1 and mGluR5 receptors are post-synaptic and mGluR2 and mGluR3 receptors are pre-synaptic.

Store at 4°C in the dark. Under these conditions, the antibodies should have a shelf life of at least twelve months, provided they remain sterile. For longer term storage, aliquot and freeze to avoid freeze-thaw of the antibody.

Chickens were immunized with three synthetic peptide / keyhole limpet hemocyanin (KLH) conjugates. These synthetic peptides corresponded to different regions of the metabotropic glutamate receptor type 1 gene product, but are shared between the human (NP_000829.2, NCBI) and mouse (NP_058672.1, NCBI) sequences. After repeated injections, immune eggs were collected, and the IgY fractions were purified from the yolks. These IgY fractions were then affinity purified using a peptide column, and the concentrations of the eluates adjusted to 300 μg/mL. Finally, equal volumes of these affinity-purified anti-peptide antibodies were mixed, and the preparation was filter-sterilized.

This anti-peptide antibody mixture was analyzed by immunohistochemistry (at a dilution of 1:2000) using fluorescein-labeled goat anti-chicken IgY (1:500 dilution, Aves Labs Cat.# F-1005) as the secondary reagent.