Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-G3BP antibody [EPR13985(B)] (ab181149)

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Western blot - Anti-G3BP antibody [EPR13985(B)] (ab181149)
  • Immunocytochemistry/ Immunofluorescence - Anti-G3BP antibody [EPR13985(B)] (ab181149)
  • Immunohistochemistry paraffin embedded sections - Anti-G3BP antibody [EPR13985(B)] (ab181149)
  • Western blot - Anti-G3BP antibody [EPR13985(B)] (ab181149)
  • Immunohistochemistry paraffin embedded sections - Anti-G3BP antibody [EPR13985(B)] (ab181149)
  • Immunocytochemistry/ Immunofluorescence - Anti-G3BP antibody [EPR13985(B)] (ab181149)
  • Flow Cytometry (Intracellular) - Anti-G3BP antibody [EPR13985(B)] (ab181149)
  • Immunoprecipitation - Anti-G3BP antibody [EPR13985(B)] (ab181149)
  • Anti-G3BP antibody [EPR13985(B)] (ab181149)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR13985(B)] to G3BP
  • Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IP
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free

Overview

  • Product name

    Anti-G3BP antibody [EPR13985(B)]
    See all G3BP primary antibodies

  • Description

    Rabbit monoclonal [EPR13985(B)] to G3BP

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IPmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • Hela, Jurkat, 293 and Ramos cell lysates; Jurkat cells, 293 cells, Human muscle tissue and Human colon tissue.

  • 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.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab181149 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
1/10000 - 1/50000. Detects a band of approximately 68 kDa (predicted molecular weight: 52 kDa).
IHC-P
1/50 - 1/100.
ICC/IF
1/250 - 1/500.
Flow Cyt
1/10.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
IP
1/20 - 1/40.
Notes
WB
1/10000 - 1/50000. Detects a band of approximately 68 kDa (predicted molecular weight: 52 kDa).
IHC-P
1/50 - 1/100.
ICC/IF
1/250 - 1/500.
Flow Cyt
1/10.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
IP
1/20 - 1/40.

Target

  • Function

    May be a regulated effector of stress granule assembly. Phosphorylation-dependent sequence-specific endoribonuclease in vitro. Cleaves exclusively between cytosine and adenine and cleaves MYC mRNA preferentially at the 3'-UTR. ATP- and magnesium-dependent helicase. Unwinds preferentially partial DNA and RNA duplexes having a 17 bp annealed portion and either a hanging 3' tail or hanging tails at both 5'- and 3'-ends. Unwinds DNA/DNA, RNA/DNA, and RNA/RNA substrates with comparable efficiency. Acts unidirectionally by moving in the 5' to 3' direction along the bound single-stranded DNA.

  • Tissue specificity

    Ubiquitous.

  • Sequence similarities

    Contains 1 NTF2 domain.
    Contains 1 RRM (RNA recognition motif) domain.

  • Domain

    The NTF2 domain mediates multimerization.

  • Post-translational
    modifications

    Phosphorylated exclusively on serine residues. Hyperphosphorylated in quiescent fibroblasts. Hypophosphorylation leads to a decrease in endoribonuclease activity (By similarity). RASA1-dependent phosphorylation of Ser-149 induces a conformational change that prevents self-association. Dephosphorylation after HRAS activation is required for stress granule assembly. Ser-149 phosphorylation induces partial nuclear localization.
    Arg-435 is dimethylated, probably to asymmetric dimethylarginine.

  • Cellular localization

    Cytoplasm. Cytoplasm > cytosol. Cell membrane. Nucleus. Cytoplasmic in proliferating cells, can be recruited to the plasma membrane in exponentially growing cells (By similarity). Cytosolic and partially nuclear in resting cells. Recruited to stress granules (SGs) upon either arsenite or high temperature treatment. Recruitment to SGs is influenced by HRAS.
  • Information by UniProt

  • Database links

    • Alternative names

      • ATP dependent DNA helicase VIII antibody
      • ATP-dependent DNA helicase VIII antibody
      • G3BP antibody
      • G3BP stress granule assembly factor 1 antibody
      • G3BP-1 antibody
      • G3bp1 antibody
      • G3BP1_HUMAN antibody
      • GAP binding protein antibody
      • GAP SH3 domain binding protein 1 antibody
      • GAP SH3 domain-binding protein 1 antibody
      • GTPase activating protein (SH3 domain) binding protein 1 antibody
      • hDH VIII antibody
      • Human DNA helicase VIII antibody
      • MGC111040 antibody
      • Ras GTPase activating protein binding protein 1 antibody
      • Ras GTPase activating protein SH3 domain binding protein antibody
      • Ras GTPase-activating protein-binding protein 1 antibody
      • RasGAP associated endoribonuclease G3BP antibody
      see all

    Images

    • Western blot - Anti-G3BP antibody [EPR13985(B)] (ab181149)
      Western blot - Anti-G3BP antibody [EPR13985(B)] (ab181149)
      All lanes : Anti-G3BP antibody [EPR13985(B)] (ab181149) at 1/10000 dilution

      Lane 1 : Wild-type A431 whole cell lysate
      Lane 2 : G3BP1 knockout A431 whole cell lysate
      Lane 3 : Jurkat whole cell lysate
      Lane 4 : HEK-293 whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 52 kDa



      Lanes 1 - 4: Merged signal (red and green). Green - ab181149 observed at 68 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab181149 was shown to specifically react with G3BP1 in wild-type A431 cells as signal was lost in G3BP1 knockout cells. Wild-type and G3BP1 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab181149 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/10000 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 1/20000 dilution for 1 hour at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence - Anti-G3BP antibody [EPR13985(B)] (ab181149)
      Immunocytochemistry/ Immunofluorescence - Anti-G3BP antibody [EPR13985(B)] (ab181149)

      Immunocytochemistry/Immunofluorescence analysis of Jurkat (Human T cell leukemia cell line from peripheral blood) labeling G3BP with Purified ab181149 at 1/500 dilution (5 µg/ml). Cells were fixed with 4% PFA and permeabilized with 0.1% tritonX-100. ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) at 1/1000 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.

    • Immunohistochemistry paraffin embedded sections - Anti-G3BP antibody [EPR13985(B)] (ab181149)
      Immunohistochemistry paraffin embedded sections - Anti-G3BP antibody [EPR13985(B)] (ab181149)

      Immunohistochemical analysis of paraffin-embedded Human colon tissue staining G3BP using ab181149 at 1/100 dilution, and prediluted HRP Polymer for Rabbit IgG as a secondary antibody with Hematoxylin counterstain.

      Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

    • Western blot - Anti-G3BP antibody [EPR13985(B)] (ab181149)
      Western blot - Anti-G3BP antibody [EPR13985(B)] (ab181149)
      All lanes : Anti-G3BP antibody [EPR13985(B)] (ab181149) at 1/20000 dilution

      Lane 1 : Hela cell lysate with NFDM/TBST
      Lane 2 : Jurkat cell lysate with NFDM/TBST
      Lane 3 : Ramos cell lysate with NFDM/TBST
      Lane 4 : 293 cell lysate with NFDM/TBST

      Lysates/proteins at 20 µg per lane.

      Blocking peptides at 5 % per lane.

      Secondary
      All lanes : Peroxidase conjugated Goat anti-Rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size: 52 kDa
      Observed band size: 68 kDa why is the actual band size different from the predicted?

    • Immunohistochemistry paraffin embedded sections - Anti-G3BP antibody [EPR13985(B)] (ab181149)
      Immunohistochemistry paraffin embedded sections - Anti-G3BP antibody [EPR13985(B)] (ab181149)

      Immunohistochemical analysis of paraffin-embedded Human muscle tissue staining G3BP using ab181149 at 1/100 dilution, and prediluted HRP Polymer for Rabbit IgG as a secondary antibody with Hematoxylin counterstain.

      Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

    • Immunocytochemistry/ Immunofluorescence - Anti-G3BP antibody [EPR13985(B)] (ab181149)
      Immunocytochemistry/ Immunofluorescence - Anti-G3BP antibody [EPR13985(B)] (ab181149)

      Immunofluorescence analysis of 4% paraformaldehyde fixed 293 cells staining G3BP using ab181149 at 1/500 dilution (red), and Goat anti rabbit IgG (Alexa Fluor® 555) at 1/200 dilution as a secondary antibody. Dapi was used for counterstaining (blue).

    • Flow Cytometry (Intracellular) - Anti-G3BP antibody [EPR13985(B)] (ab181149)
      Flow Cytometry (Intracellular) - Anti-G3BP antibody [EPR13985(B)] (ab181149)

      Flow cytometric analysis of 2% paraformaldehyde fixed Jurkat cells staining G3BP using ab181149 at 1/10 dilution, and FITC conjugated Goat anti rabbit IgG at 1/150 dilution as a secondary antibody (red curve). Rabbit monoclonal IgG was the negative control (green curve).

       

    • Immunoprecipitation - Anti-G3BP antibody [EPR13985(B)] (ab181149)
      Immunoprecipitation - Anti-G3BP antibody [EPR13985(B)] (ab181149)

      Western blot analysis of G3BP in immunoprecipitation pellets from Ramos cell lysate, using ab181149 at a 1/50 dilution. Anti-Rabbit IgG (HRP) specific to the non-reduced form of IgG was used as a secondary antibody at 1/1000 dilution. Blocking/dilution buffer and concentration: 5% NFDM/TBST

    • Anti-G3BP antibody [EPR13985(B)] (ab181149)
      Anti-G3BP antibody [EPR13985(B)] (ab181149)

    References (6)

    Publishing research using ab181149? Please let us know so that we can cite the reference in this datasheet.

    ab181149 has been referenced in 6 publications.

    • Christ W  et al. Puumala and Andes Orthohantaviruses Cause Transient Protein Kinase R-Dependent Formation of Stress Granules. J Virol 94:N/A (2020). PubMed: 31723021
    • Fernandes N  et al. Stress Granule Assembly Can Facilitate but Is Not Required for TDP-43 Cytoplasmic Aggregation. Biomolecules 10:N/A (2020). PubMed: 32992901
    • Sonobe Y  et al. Translation of dipeptide repeat proteins from the C9ORF72 expanded repeat is associated with cellular stress. Neurobiol Dis 116:155-165 (2018). PubMed: 29792928
    • Zhang K  et al. Stress Granule Assembly Disrupts Nucleocytoplasmic Transport. Cell 173:958-971.e17 (2018). PubMed: 29628143
    • Tsai WC  et al. Histone arginine demethylase JMJD6 is linked to stress granule assembly through demethylation of the stress granule-nucleating protein G3BP1. J Biol Chem 292:18886-18896 (2017). PubMed: 28972166
    • Tsai WC  et al. Arginine Demethylation of G3BP1 Promotes Stress Granule Assembly. J Biol Chem 291:22671-22685 (2016). PubMed: 27601476

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