Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-GNAQ antibody [EPR20978] (ab210004)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20978] to GNAQ
  • Suitable for: IP, Flow Cyt, IHC-P, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-GNAQ antibody [EPR20978]
    See all GNAQ primary antibodies

  • Description

    Rabbit monoclonal [EPR20978] to GNAQ

  • Host species

    Rabbit

  • Specificity

    The sequence homology between GNAQ and three other related Ga family members: GNA11, GNA14 and GNA15 is 90.3%, 79.9% and 53.5%, respectively.

  • Tested applications

    Suitable for: IP, Flow Cyt, IHC-P, WBmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant full length protein within Human GNAQ aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: P50148

  • Positive control

    • WB: HEK-293T, HeLa, A549, Jurkat, RAW 264.7, PC-12 and C6 whole cell lysates; Human stomach and fetal pancreas lysates; Mouse lung and kidney lysates; Rat lung lysate; His-tagged human GNAQ (aa1-359) recombinant protein. IHC-P: Human mammary gland and papillary thyroid carcinoma tissues. Flow Cyt: HEK-293T cells. IP: A549 whole cell lysate.

  • 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
    For more information see here.

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

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Applications

Our Abpromise guarantee covers the use of ab210004 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
IP 1/30.
Flow Cyt 1/50.
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

This antibody is not recommended for mouse and rat species in IHC application. Under our experiment conditions this antibody showed weak or no staining on mouse and rat tissues.
WB 1/1000. Detects a band of approximately 42 kDa (predicted molecular weight: 42 kDa).

Target

  • Function

    Guanine nucleotide-binding proteins (G proteins) are involved as modulators or transducers in various transmembrane signaling systems.

  • Tissue specificity

    Predominantly expressed in ovary, prostate, testis and colon.

  • Sequence similarities

    Belongs to the G-alpha family. G(q) subfamily.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • CMC1 antibody
    • G alpha Q antibody
    • G protein alpha Q antibody
    • G protein subunit alpha q antibody
    • G-ALPHA-q antibody
    • GAQ antibody
    • GNAQ antibody
    • GNAQ_HUMAN antibody
    • guanine nucleotide binding protein (G protein), q polypeptide antibody
    • Guanine nucleotide binding protein alpha q antibody
    • guanine nucleotide binding protein G protein q polypeptide antibody
    • Guanine nucleotide binding protein G q subunit alpha antibody
    • Guanine nucleotide-binding protein alpha-q antibody
    • Guanine nucleotide-binding protein G(q) subunit alpha antibody
    • SWS antibody
    see all

Images

  • Western blot - Anti-GNAQ antibody [EPR20978] (ab210004)
    Western blot - Anti-GNAQ antibody [EPR20978] (ab210004)

    Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: GNAQ knockout HAP1 whole cell lysate (20 µg)

    Lanes 1 - 2: Merged signal (red and green). Green - ab210004 observed at 42 kDa. Red - loading control, ab18058, observed at 130 kDa.

    ab210004 was shown to specifically react with GNAQ in wild-type HAP1 cells as signal was lost in GNAQ knockout cells. Wild-type and GNAQ knockout samples were subjected to SDS-PAGE. ab210004 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-GNAQ antibody [EPR20978] (ab210004)
    Western blot - Anti-GNAQ antibody [EPR20978] (ab210004)
    All lanes : Anti-GNAQ antibody [EPR20978] (ab210004) at 1/1000 dilution

    Lane 1 : His-tagged human GNAQ (aa1-359) recombinant protein
    Lane 2 : His-tagged human GNA11 (aa1-359) recombinant protein

    Lysates/proteins at 0.01 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Predicted band size: 42 kDa
    Observed band size: 42 kDa


    Exposure time: 1 second


    Blocking and dilution buffer: 5% NFDM/TBST 

  • Western blot - Anti-GNAQ antibody [EPR20978] (ab210004)
    Western blot - Anti-GNAQ antibody [EPR20978] (ab210004)
    Lanes 1-4 : Anti-GNAQ antibody [EPR20978] (ab210004) at 1/2000 dilution
    Lanes 5-7 : Anti-GNAQ antibody [EPR20978] (ab210004) at 1/1000 dilution

    Lane 1 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
    Lane 2 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
    Lane 3 : A549 (human lung carcinoma cell line) whole cell lysate at 20 µg
    Lane 4 : Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg
    Lane 5 : C6 (rat glial tumor cell line) whole cell lysate at 10 µg
    Lane 6 : RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg
    Lane 7 : PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate at 10 µg

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Predicted band size: 42 kDa
    Observed band size: 42 kDa



     

    Exposure time : Lanes 1-3 & 5-7: 3 minutes; Lane 4: 3 seconds.

    Blocking and dilution buffer: 5% NFDM/TBST

  • Western blot - Anti-GNAQ antibody [EPR20978] (ab210004)
    Western blot - Anti-GNAQ antibody [EPR20978] (ab210004)
    Lanes 1-4 : Anti-GNAQ antibody [EPR20978] (ab210004) at 1/2000 dilution
    Lane 5 : Anti-GNAQ antibody [EPR20978] (ab210004) at 1/1000 dilution

    Lane 1 : Human fetal pancreas lysate at 20 µg
    Lane 2 : Mouse lung lysate at 20 µg
    Lane 3 : Mouse kidney lysate at 20 µg
    Lane 4 : Rat lung lysate at 20 µg
    Lane 5 : Human stomach lysate at 10 µg

    Secondary
    Lanes 1-4 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
    Lane 5 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution

    Developed using the ECL technique.

    Predicted band size: 42 kDa
    Observed band size: 42 kDa



     

    Exposure time : Lanes 2-4: 3 minutes; Lanes 1 & 5: 15 seconds.

    Blocking and dilution buffer: 5% NFDM/TBST

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GNAQ antibody [EPR20978] (ab210004)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GNAQ antibody [EPR20978] (ab210004)

    Immunohistochemical analysis of paraffin-embedded human mammary gland tissue labeling GNAQ with ab210004 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous and cytoplasmic staining on human mammary gland is observed (PMID: 22421440; PMID: 1946421; PMID: 18799799). Counter stained with hematoxylin.


    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

     

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GNAQ antibody [EPR20978] (ab210004)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GNAQ antibody [EPR20978] (ab210004)

    Immunohistochemical analysis of paraffin-embedded human papillary thyroid carcinoma tissue labeling GNAQ with ab210004 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous and cytoplasmic staining on human mammary gland is observed (PMID: 22421440; PMID: 1946421; PMID: 18799799). Counter stained with hematoxylin.


    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Flow Cytometry - Anti-GNAQ antibody [EPR20978] (ab210004)
    Flow Cytometry - Anti-GNAQ antibody [EPR20978] (ab210004)

    Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methonol-permeabilized HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cell line labeling GNAQ with ab210004 at 1/50 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.

  • Immunoprecipitation - Anti-GNAQ antibody [EPR20978] (ab210004)
    Immunoprecipitation - Anti-GNAQ antibody [EPR20978] (ab210004)

    GNAQ was immunoprecipitated from 0.35 mg of A549 (human lung carcinoma cell line) whole cell lysate with ab210004 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab210004 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: A549 whole cell lysate 10 μg (Input).

    Lane 2: ab210004 IP in A549 whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab210004 in A549 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 minutes.

  • Anti-GNAQ antibody [EPR20978] (ab210004)
    Anti-GNAQ antibody [EPR20978] (ab210004)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (1)

Publishing research using ab210004? Please let us know so that we can cite the reference in this datasheet.

ab210004 has been referenced in 1 publication.

  • Govatati S  et al. NFATc1-E2F1-LMCD1-Mediated IL-33 Expression by Thrombin Is Required for Injury-Induced Neointima Formation. Arterioscler Thromb Vasc Biol 39:1212-1226 (2019). PubMed: 31043075

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