Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-GNAQ antibody [EPR20978] - BSA and Azide free (ab225744)

Overview

  • Product name

    Anti-GNAQ antibody [EPR20978] - BSA and Azide free
    See all GNAQ primary antibodies
  • Description

    Rabbit monoclonal [EPR20978] to GNAQ - BSA and Azide free
  • 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: WB, IHC-P, Flow Cyt, IPmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant full length protein within Human GNAQ aa 1-400. The exact sequence is proprietary.
    Database link: P50148

  • Positive control

    • IHC-P: Human mammary gland tissue.
  • General notes

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

    ab225744 is a PBS-only buffer format of ab210004. Please refer to ab210004 for recommended dilutions, protocols, and image data.

     

    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 ab225744 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. Detects a band of approximately 42 kDa (predicted molecular weight: 42 kDa).
IHC-P Use at an assay dependent concentration. 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.

Flow Cyt Use at an assay dependent concentration.
IP Use at an assay dependent concentration.

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

  • 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

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

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : GNAQ knockout HAP1 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 42 kDa



    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.

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

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

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

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab210004).

References

ab225744 has not yet been referenced specifically in any publications.

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