Overview

  • Product nameAnti-GNAQ antibody
    See all GNAQ primary antibodies
  • Description
    Rabbit polyclonal to GNAQ
  • SpecificityThe immunogen has 66% homology with GNA11
  • Tested applicationsSuitable for: IHC-P, WB, ICC/IF, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Dog, Pig, Xenopus laevis
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human GNAQ.

    (Peptide available as ab86404.)

  • Positive control
    • This antibody gave a positive signal in the following lysates: HepG2 Whole Cell; Mouse Pancreas Tissue; Mouse Kidney Tissue; Rat Liver Tissue. This antibody gave a positive result in IHC in the following FFPE tissue - Human normal lung

Properties

Applications

Our Abpromise guarantee covers the use of ab75825 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-P Use a concentration of 1 - 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 42 kDa (predicted molecular weight: 42 kDa).
ICC/IF Use a concentration of 5 µg/ml.
IP Use at an assay dependent concentration.

Target

Anti-GNAQ antibody images

  • IHC image of GNAQ staining in Human normal lung formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab75825, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • All lanes : Anti-GNAQ antibody (ab75825) at 1 µg/ml

    Lane 1 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 2 : Pancreas (Mouse) Tissue Lysate
    Lane 3 : Kidney (Mouse) Tissue Lysate
    Lane 4 : Liver (Rat) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP)
    Developed using the ECL technique

    Predicted band size : 42 kDa
    Observed band size : 42 kDa
    Additional bands at : 70 kDa. We are unsure as to the identity of these extra bands.
  • GNAQ was immunoprecipitated using 0.5mg HepG2 whole cell extract, 5µg of Rabbit polyclonal to GNAQ and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, HepG2 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab75825.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 42kDa: GNAQ.
  • IHC image of G protein alpha q staining in human colon formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab75825, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
  • ICC/IF image of ab75825 stained Hek293 cells. The cells were 4% paraformaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab75825, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% paraformaldehyde fixed (10 min) HepG2 cells at 5µg/ml, and in 100% methanol fixed (5 min) Hek293, HepG2 and MCF7 cells at 5µg/ml.

References for Anti-GNAQ antibody (ab75825)

This product has been referenced in:
  • Mann KM  et al. Sleeping Beauty mutagenesis reveals cooperating mutations and pathways in pancreatic adenocarcinoma. Proc Natl Acad Sci U S A 109:5934-41 (2012). IHC-P ; Mouse . Read more (PubMed: 22421440) »
  • Mak S  et al. Differential expression of genes in the calcium-signaling pathway underlies lesion development in the LDb mouse model of atherosclerosis. Atherosclerosis 213:40-51 (2010). Read more (PubMed: 20667539) »

See all 2 Publications for this product

Product Wall

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (human uveal melanoma)
Permeabilization Yes - 0.1% saponin
Specification human uveal melanoma
Blocking step BSA as blocking agent for 1 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative Formaldehyde
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Submitted May 11 2016

Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (Tumor)
Specification Tumor
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative Paraformaldehyde
Username

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Verified customer

Submitted Nov 30 2015

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"