Recombinant Anti-Metabotropic Glutamate Receptor 5 antibody [EPR2425Y] (ab76316)

ab76316 staining Metabotropic Glutmate Receptor 5 in rat hippocampal neurons by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with 0.2% Triton X-100 and blocked with 5% serum for 20 minutes at 22°C. Samples were incubated with primary antibody (1/200) for 24 hours at 4°C. An Alexa Fluor® 594-conjugated chicken anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody.

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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^oC

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.

Array tomography of adult mouse neocortex using Anti-Metabotropic Glutamate Receptor 5 antibody [EPR2425Y] (ab76316) (green), VGluT1 (red) and DAPI (blue).

Tissue was fixed with 4% paraformaldehyde before being embedded in LRWhite resin. Ultra-thin sections (70 nm) were incubated with Anti-Metabotropic Glutamate Receptor 5 antibody [EPR2425Y] (ab76316) at a concentration of 1:13 overnight at 4^oC. Secondary antibodies were applied for 30 minutes at room temperature.

Image courtesy of Dr. Kristina Micheva, Stanford University School of Medicine.

See full array tomography protocol.

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.

See Abreview