Immunohistochemical analysis of Paraffin-embedded rat cerebrum tissue sections labeling Metabotropic Glutamate Receptor 5 with ab76316 at 1/400 dilution followed by Goat Anti-Rabbit IgG H&L (HRP Polymer) secondary antibody. Sections were counterstained with Hematoxylin. Antigen retrieval was heat mediated using ab93684 (Tris/EDTA buffer, pH 9.0).

Positive staining on rat cerebrum.

ab76316 staining Metabotropic Glutmate Receptor 5 in rat caudate putamen by immunohistochemistry. Tissue was fixed with formaldehyde and citrate-mediated antigen retrieval was performed. Samples were blocked with 1% BSA for 10 minutes at 21°C, before incubation with the primary antibody (1/2000) for 2 hours at 21°C. A biotin conjugated goatanti-rabbit IgG secondary was used at 1/250.

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ab76316 staining Metabotropic Glutmate Receptor 5 in mouse caudate putamen/ Corpus callosum by immunohistochemistry (frozen sections). Tissue was fixed with formaldehyde and samples were blocked with 1% BSA for 10 minutes at 21°C, before incubation with the primary antibody (1/2000) for 16 hours at 21°C. An alexa fluor® 594 conjugated goat anti-rabbit IgG secondary was used at 1/500.

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Postembedding immunogold labeling of mouse neocortex using Anti-Metabotropic Glutamate Receptor 5 antibody [EPR2425Y] (ab76316). The tissue was embedded in Lowicryl HM20 resin. 60 nm sections were then cut and mounted on nickel mesh grids before undergoing antigen retrieval for 15 minutes in 0.01 M citrate buffer, pH 6 at 60°C.

The sections were then blocked in 1% BSA/TBSN pH 7.6 and incubated overnight at room temperature with Anti-Metabotropic Glutamate Receptor 5 antibody [EPR2425Y] (ab76316) at 1:250. Sections were washed twice in TBSN pH 7.6, treated with 1% normal donkey serum/TBSN pH 8.2 for 30 minutes, before being incubated with donkey anti-rabbit IgG-Au 10-20 nm at 1:20 for two hours at room temperature.

Sections were then washed in TBSN pH 8.2, followed by water, before undergoing post-staining with 1% uranyl acetate and Sato’s lead. They were then air dried before being transferred to an oven for 30 minutes at 60°C.

In these images you can see gold immunoparticles on the postsynaptic density of synapses.

(TBSN = 0.02M TRIS buffered saline (0.3 N, pH 7.6 or 8.2) with 0.005% Tergitol NP-10)

Flow cytometric analysis of permeabilized SH-SY5Y cells using ab76316 (red) at 1/20 or a rabbit IgG (ab172730) as a negative control (green). The cells were permeabilized with 2% PFA and a goat anti-rabbit IgG FITC was used as the secondary at 1/150.

ab76316 staining Metabotropic Glutmate Receptor 5 in mouse caudate putamen/ Corpus callosum by immunohistochemistry. Tissue was fixed with formaldehyde and citrate-mediated antigen retrieval was performed. Samples were blocked with 1% BSA for 10 minutes at 21°C, before incubation with the primary antibody (1/1000) for 2 hours at 21°C. A biotin conjugated goatanti-rabbit IgG secondary was used at 1/250.

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