Recombinant
RabMAb

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

Overview

  • 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
  • Tested applications
    Suitable for: IHC-R, WB, IHC-P, Flow Cyt, Electron Microscopy, ICC/IF, IHC-Frmore details
    Unsuitable for: IP
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Metabotropic Glutamate Receptor 5 aa 1150 to the C-terminus (C terminal). The exact sequence is proprietary.
    (Peptide available as ab139974)

  • Positive control
    • WB: Mouse brain lysate. ICC/IF: Rat hippocampal neurons. IHC-P: Rat caudate putamen tissue; mouse caudate putamen tissue; human brain tissue. IHC-Fr: Mouse caudate ptamen tissue. IHC (resin): Mouse neocortex tissue. EM: Mouse neocortex tissue. Flow cyt: SH-SY5Y cells.
  • General notes

    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 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
IHC-R 1/13.
WB 1/5000 - 1/10000. Predicted molecular weight: 132 kDa.Can be blocked with Metabotropic Glutamate Receptor 5 peptide (ab139974).
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.

Flow Cyt 1/10.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Electron Microscopy Use at an assay dependent concentration.
ICC/IF 1/100 - 1/250.
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

    • 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)

    • 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.

      See Abreview

    • 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.

      See Abreview

    • 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.

      See Abreview

    • 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)

    • 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 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.

    • 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

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

    References

    This product has been referenced in:
    • Huang M  et al. Metabotropic glutamate receptor 5 signalling induced NMDA receptor subunits alterations during the development of morphine-induced antinociceptive tolerance in mouse cortex. Biomed Pharmacother 110:717-726 (2019). Read more (PubMed: 30553198) »
    • Liu YW  et al. Transplantation with mGluR5 deficiency bone marrow displays antidepressant-like effect in C57BL/6J mice. Brain Behav Immun N/A:N/A (2019). Read more (PubMed: 30682501) »
    See all 13 Publications for this product

    Customer reviews and Q&As

    1-9 of 9 Abreviews or Q&A

    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Human Tissue sections (Skin)
    Permeabilization
    No
    Specification
    Skin
    Blocking step
    Serum as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 10% · Temperature: 20°C
    Fixative
    Acetone

    Abcam user community

    Verified customer

    Submitted May 10 2019

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Monkey Cell (COS7)
    Permeabilization
    Yes - 0.2%Triton
    Specification
    COS7
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 22°C
    Fixative
    Formaldehyde

    Abcam user community

    Verified customer

    Submitted Oct 07 2015

    Application
    Immunohistochemistry (Frozen sections)
    Blocking step
    BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
    Sample
    Mouse Tissue sections (Brain)
    Specification
    Brain
    Permeabilization
    No
    Fixative
    Formaldehyde

    Mr. Carl Hobbs

    Verified customer

    Submitted Aug 11 2014

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step
    BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citric acid
    Sample
    Rat Tissue sections (Brain)
    Specification
    Brain
    Permeabilization
    No
    Fixative
    Formaldehyde

    Mr. Carl Hobbs

    Verified customer

    Submitted Aug 11 2014

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step
    BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citric acid
    Sample
    Mouse Tissue sections (Brain)
    Specification
    Brain
    Permeabilization
    No
    Fixative
    Formaldehyde

    Mr. Carl Hobbs

    Verified customer

    Submitted Aug 11 2014

    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    Serum as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 22°C
    Sample
    Rat Cell (rat hippocampal neuron)
    Specification
    rat hippocampal neuron
    Permeabilization
    Yes - 0.2%Triton
    Fixative
    Formaldehyde

    Abcam user community

    Verified customer

    Submitted Apr 13 2014

    Answer



    I am happy to confirm that we have the suitable blocking peptide available in our catalog under the product number ab139974

    https://www.abcam.com/index.html?datasheet=139974 (or use the following: https://www.abcam.com/index.html?datasheet=139974).

    Read More

    Answer

    Thank you for your reply.

    The blocking peptide for the Anti-Metabotropic Glutamate Receptor 5 antibody [EPR2425Y] (ab76316) is now available to purchase under the catalogue number ab139974. The details of this product can be viewed from the following link:

    https://www.abcam.com/Metabotropic-Glutamate-Receptor-5-peptide-ab139974.html

    If you have any further questions, please do not hesitate to ask.

    Read More

    Answer

    Sorry for the delay in getting back to you.

    I have had confirmation that we do have the blocking peptide available for the rabbit monoclonal anti-Metabotropic Glutamate Receptor 5 antibody [EPR2425Y] (ab76316). If you would like for me to add this to the catalogue for your customer please do let me know.

    I have now arranged for the replacement antibody ab76316 to be sent to you. This is on the order number1111586. Please bear in mind that the item may be moved from this order before shipping through consolidation of the orders. If you would like to track the item please quote "FOCR CCE4186704" to any of our customer service representatives.

    I hope this replacement works well for the customer. If they continue to have any problems or if I can be of any further assistance, please do letme know.

    Read More

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