Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-GNAQ antibody [EPR20978] (ab210004)

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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

    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 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.

References

ab210004 has not yet been referenced specifically in any publications.

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

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