Lane 1 : mGluR5 antibody at 1/600 dilutionLanes 2 - 4 : Anti-Metabotropic Glutamate Receptor 5 (phospho S996) antibody (ab16653) at 3 µg/mlLane 1 : mouse hippocampal lysateLane 2 : mouse hippocampal lysateLane 3 : mouse hippocampal lysate with Rat Metabotropic Glutamate Receptor 5 (phospho S996) peptide (ab17573) at 3 µg/mlLane 4 : mouse hippocampal lysate with Rat Metabotropic Glutamate Receptor 5 peptide (ab17575) at 3 µg/mlLysates/proteins at 10 µg per lane.Performed under reducing conditions.
ICC/IF image of ab16653 stained rat PC12 cells. The cells were methanol fixed (5 min), permabilised in PBS-T (20 min) and incubated with the antibody (ab16653, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
IHC-FoFr image of Metaobotropic Glutamate Receptor 5 (pS996) staining on rat brain sections using ab16653 (1:300). The sections used came from animals perfused fixed with Paraformaldehyde 4% with 15% of a solution of saturated picric acid, in phosphate buffer 0.1M. Following postfixation in the same fixative overnight, the brains were cryoprotected in sucrose 30% overnight. Brains were then cut using a cryostat and the immunostainings were performed using the ‘free floating’ technique.See Abreview