Recombinant Anti-Metabotropic Glutamate Receptor 5 antibody [EPR2425Y] (ab76316)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR2425Y] to Metabotropic Glutamate Receptor 5
- Suitable for: Electron Microscopy, Flow Cyt (Intra), WB, IHC-P, IHC-Fr
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Metabotropic Glutamate Receptor 5 antibody [EPR2425Y]
See all Metabotropic Glutamate Receptor 5 primary antibodies -
Description
Rabbit monoclonal [EPR2425Y] to Metabotropic Glutamate Receptor 5 -
Host species
Rabbit -
Specificity
The Human species recommendation is based on the WB results. This antibody has been tested for IHC-P and IHC-Fr in Human samples and we obtain positive signal only in IHC-Fr. We do not recommend this antibody for IHC-P in Human samples.
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Tested applications
Suitable for: Electron Microscopy, Flow Cyt (Intra), WB, IHC-P, IHC-Frmore details
Unsuitable for: IP -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Mouse brain lysate and Human hippocampus tissue. IHC-P: Rat caudate putamen and cerebrum tissue; mouse caudate putamen tissue and cerebrum IHC-Fr: Mouse caudate ptamen tissue. IHC (resin): Mouse neocortex tissue. EM: Mouse neocortex tissue. Flow Cyt (intra): SH-SY5Y cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR2425Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- HRP Anti-Metabotropic Glutamate Receptor 5 antibody [EPR2425Y] (ab196482)
- Alexa Fluor® 647 Anti-Metabotropic Glutamate Receptor 5 antibody [EPR2425Y] (ab196929)
- Anti-Metabotropic Glutamate Receptor 5 antibody [EPR2425Y] - Low endotoxin, Azide free (ab219374)
- Anti-Metabotropic Glutamate Receptor 5 antibody [EPR2425Y] - BSA and Azide free (ab256589)
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab76316 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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Electron Microscopy |
Use at an assay dependent concentration.
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Flow Cyt (Intra) |
1/10.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB |
1/5000 - 1/10000. Predicted molecular weight: 132 kDa.
Please do not boil the sample before loading to the gel. |
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IHC-P | (2) |
1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Use of HRP-conjugated or polymerized HRP secondary antibodies, stronger signals have been found using the polymerized HRP secondary. We only recommend mouse and rat species for IHC-P. We do not guarantee IHC for Human species. |
IHC-Fr | (2) |
Use a concentration of 1 µg/ml.
This antibody has been tested for IHC-P and IHC-Fr in Human samples and we obtain positive signal only in IHC-Fr. We do not recommend this antibody for IHC-P in Human samples. |
Notes |
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Electron Microscopy
Use at an assay dependent concentration. |
Flow Cyt (Intra)
1/10. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/5000 - 1/10000. Predicted molecular weight: 132 kDa. Please do not boil the sample before loading to the gel. |
IHC-P
1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. Use of HRP-conjugated or polymerized HRP secondary antibodies, stronger signals have been found using the polymerized HRP secondary. We only recommend mouse and rat species for IHC-P. We do not guarantee IHC for Human species. |
IHC-Fr
Use a concentration of 1 µg/ml. This antibody has been tested for IHC-P and IHC-Fr in Human samples and we obtain positive signal only in IHC-Fr. We do not recommend this antibody for IHC-P in Human samples. |
Target
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Function
G-protein coupled receptor for glutamate. Ligand binding causes a conformation change that triggers signaling via guanine nucleotide-binding proteins (G proteins) and modulates the activity of down-stream effectors. Signaling activates a phosphatidylinositol-calcium second messenger system and generates a calcium-activated chloride current. Plays an important role in the regulation of synaptic plasticity and the modulation of the neural network activity. -
Sequence similarities
Belongs to the G-protein coupled receptor 3 family. -
Cellular localization
Cell membrane. - Information by UniProt
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Database links
- Entrez Gene: 2915 Human
- Entrez Gene: 108071 Mouse
- Entrez Gene: 24418 Rat
- Omim: 604102 Human
- SwissProt: P41594 Human
- SwissProt: Q3UVX5 Mouse
- SwissProt: P31424 Rat
- Unigene: 147361 Human
see all -
Alternative names
- Glutamate receptor metabotropic 5 antibody
- GPRC1E antibody
- Grm5 antibody
see all
Images
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Immunohistochemistry (Frozen sections) - Anti-Metabotropic Glutamate Receptor 5 antibody [EPR2425Y] (ab76316)
IHC image of Metabotropic Glutamate Receptor 5 staining in a section of frozen normal human hippocampus* performed on a Leica BOND™ system using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab76316, 1 µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre -
All lanes : Anti-Metabotropic Glutamate Receptor 5 antibody [EPR2425Y] (ab76316) at 1/10000 dilution
Lane 1 : Human spinal cord tissue lysate was not boiled before loading to the gel
Lane 2 : Human hippocampus tissue lysate was not boiled before loading to the gel
Lane 3 : Human liver tissue lysate was not boiled before loading to the gel
Lane 4 : Human spinal cord tissue lysate was boiled before loading to the gel
Lane 5 : Human hippocampus tissue lysate was boiled before loading to the gel
Lane 6 : Human liver tissue lysate was boiled before loading to the gel
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 132 kDa
Observed band size: 140 kDa why is the actual band size different from the predicted?
Additional bands at: 280 kDa (possible dimer)Buffer: 5% NFDM /TBST
Please do not boil the sample before loading to the gel.
GRM5 is a brain enriched protein, can be observed in most of the brain areas but low in spinal cord (PMID: 12783878).
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Anti-Metabotropic Glutamate Receptor 5 antibody [EPR2425Y] (ab76316) at 1/10000 dilution + mouse brain lysate at 10 µg
Secondary
Goat anti-rabbit-HRP
Developed using the ECL technique.
Predicted band size: 132 kDa
Observed band size: 140-150 kDa why is the actual band size different from the predicted?
Additional bands at: >250 kDa (possible dimer) -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Metabotropic Glutamate Receptor 5 antibody [EPR2425Y] (ab76316)Immunohistochemical analysis of Paraffin-embedded mouse 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 mouse cerebrum.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Metabotropic Glutamate Receptor 5 antibody [EPR2425Y] (ab76316)
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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Metabotropic Glutamate Receptor 5 antibody [EPR2425Y] (ab76316)Image courtesy of Carl Hobbs, King College London, U.K.
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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Immunohistochemistry (Frozen sections) - Anti-Metabotropic Glutamate Receptor 5 antibody [EPR2425Y] (ab76316)Image courtesy of Carl Hobbs, King College London, U.K.
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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Electron Microscopy - Anti-Metabotropic Glutamate Receptor 5 antibody [EPR2425Y] (ab76316)Images courtesy of Professor Richard Weinberg, UNC School of Medicine
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)
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Flow Cytometry (Intracellular) - Anti-Metabotropic Glutamate Receptor 5 antibody [EPR2425Y] (ab76316)
Intracellular 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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Metabotropic Glutamate Receptor 5 antibody [EPR2425Y] (ab76316)Image courtesy of Carl Hobbs, King College London, U.K.
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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (31)
ab76316 has been referenced in 31 publications.
- Huang M et al. Deficiency of SHANK3 isoforms impairs thermal hyperalgesia and dysregulates the expression of postsynaptic proteins in the spinal cord. Neurosci Res 163:26-33 (2021). PubMed: 32147472
- Chiprés-Tinajero GA et al. Increased immunoreactivity of glutamate receptors, neuronal nuclear protein and glial fibrillary acidic protein in the hippocampus of epileptic rats with fast ripple activity. Exp Brain Res 239:2015-2024 (2021). PubMed: 33909110
- Elhussiny MEA et al. Modulation by chronic stress and ketamine of ionotropic AMPA/NMDA and metabotropic glutamate receptors in the rat hippocampus. Prog Neuropsychopharmacol Biol Psychiatry 104:110033 (2021). PubMed: 32640261
- Peng K et al. Mechanisms Underlying Enhancement of Spontaneous Glutamate Release by Group I mGluRs at a Central Auditory Synapse. J Neurosci 40:7027-7042 (2020). PubMed: 32801152
- Wang Y et al. mGluR5 mediates ketamine antidepressant response in susceptible rats exposed to prenatal stress. J Affect Disord 272:398-408 (2020). PubMed: 32553383