Recombinant
RabMAb

Recombinant Anti-Metabotropic Glutamate Receptor 5 antibody [EPR2425Y] - Low endotoxin, Azide free (ab219374)

Overview

  • Product name
    Anti-Metabotropic Glutamate Receptor 5 antibody [EPR2425Y] - Low endotoxin, Azide free
    See all Metabotropic Glutamate Receptor 5 primary antibodies
  • Description
    Rabbit monoclonal [EPR2425Y] to Metabotropic Glutamate Receptor 5 - Low endotoxin, Azide free
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, Flow Cyt, ICC/IF, Electron Microscopy, IHC-R, IHC-Frmore details
    Unsuitable for: IP
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Metabotropic Glutamate Receptor 5 aa 1150 to the C-terminus (C terminal).
    (Peptide available as ab139974)

  • Positive control
    • Human brain tissue, mouse brain lysate.
  • General notes

    The formulation and the concentration of this product is compatible for metal-conjugation for mass cytometry (CyTOF®).

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab219374 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
WB Use at an assay dependent concentration. Predicted molecular weight: 132 kDa.Can be blocked with Metabotropic Glutamate Receptor 5 peptide (ab139974).
IHC-P Use at an assay dependent concentration. 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.

Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF Use at an assay dependent concentration.
Electron Microscopy Use at an assay dependent concentration.
IHC-R Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for IP.
  • Target

    • 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
    • Database links
    • Alternative names
      • Glutamate receptor metabotropic 5 antibody
      • GPRC1E antibody
      • Grm5 antibody
      • GRM5_HUMAN antibody
      • Metabotropic glutamate receptor 5 antibody
      • Metabotropic glutamate receptor 5 variant F antibody
      • Metabotropic glutamate receptor 5 variant G antibody
      • Metabotropic glutamate receptor 5 variant H antibody
      • mGlu5 antibody
      • mGluR5 antibody
      • PPP1R86 antibody
      • Protein phosphatase 1 regulatory subunit 86 antibody
      see all

    Images

    • 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76316).

    • 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 4oC. 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76316).

    • 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 60oC

      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)

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76316).

    • 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76316).

    • 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76316).

    • 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76316).

    • Human brain tissue with ab76316 at 1/250 dilution.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76316).

    References

    This product has been referenced in:
    • Lee KW  et al. Alteration by p11 of mGluR5 localization regulates depression-like behaviors. Mol Psychiatry 20:1546-56 (2015). Read more (PubMed: 26370144) »
    • Golubeva AV  et al. The mouse cyclophosphamide model of bladder pain syndrome: tissue characterization, immune profiling, and relationship to metabotropic glutamate receptors. Physiol Rep 2:e00260 (2014). WB ; Mouse . Read more (PubMed: 24760514) »
    See all 5 Publications for this product

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