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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-G3BP antibody [EPR13986(B)] (ab181150)

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  • Certificate of Compliance
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Western blot - Anti-G3BP antibody [EPR13986(B)] (ab181150)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-G3BP antibody [EPR13986(B)] (ab181150)
  • Western blot - Anti-G3BP antibody [EPR13986(B)] (ab181150)
  • Immunocytochemistry/ Immunofluorescence - Anti-G3BP antibody [EPR13986(B)] (ab181150)
  • Western blot - Anti-G3BP antibody [EPR13986(B)] (ab181150)
  • Immunoprecipitation - Anti-G3BP antibody [EPR13986(B)] (ab181150)
  • Flow Cytometry - Anti-G3BP antibody [EPR13986(B)] (ab181150)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-G3BP antibody [EPR13986(B)] (ab181150)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-G3BP antibody [EPR13986(B)] (ab181150)
  • Immunocytochemistry/ Immunofluorescence - Anti-G3BP antibody [EPR13986(B)] (ab181150)
  • Western blot - Anti-G3BP antibody [EPR13986(B)] (ab181150)
  • Immunohistochemistry paraffin embedded sections - Anti-G3BP antibody [EPR13986(B)] (ab181150)
  • Immunoprecipitation - Anti-G3BP antibody [EPR13986(B)] (ab181150)
  • Anti-G3BP antibody [EPR13986(B)] (ab181150)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR13986(B)] to G3BP
  • Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

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Overview

  • Product name

    Anti-G3BP antibody [EPR13986(B)]
    See all G3BP primary antibodies
  • Description

    Rabbit monoclonal [EPR13986(B)] to G3BP
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human G3BP aa 400 to the C-terminus. The exact sequence is proprietary.
    Database link: Q13283

  • Positive control

    • WB: NIH/3T3, PC-12, Hela, Jurkat, A431, Ramos and 293 cell lysates IHC-P: Human hepatocellular cancer tissue, Human colon tissue, mouse and rat kidney tissue. ICC/IF: HeLa and 293 cells Flow Cyt: HeLa cells IP: Ramos 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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR13986(B)
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • DNA / Nucleotides
    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • RNA Processing
    • Other

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-G3BP antibody [EPR13986(B)] (ab214946)
    • Alexa Fluor® 647 Anti-G3BP antibody [EPR13986(B)] (ab215944)
    • Alexa Fluor® 594 Anti-G3BP antibody [EPR13986(B)] (ab217225)
    • Alexa Fluor® 555 Anti-G3BP antibody [EPR13986(B)] (ab217729)
    • Alexa Fluor® 568 Anti-G3BP antibody [EPR13986(B)] (ab220524)
    • Anti-G3BP antibody [EPR13986(B)] - BSA and Azide free (ab240247)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • HeLa whole cell lysate (ab150035)
    • HeLa whole cell lysate (ab29545)
    • Ramos whole cell lysate (ab3955)
    • Jurkat whole cell lysate (ab7899)
  • Recombinant Protein

    • Recombinant Human G3BP protein (ab103304)
  • Related Products

    • Recombinant Human G3BP protein (ab103304)

Applications

Our Abpromise guarantee covers the use of ab181150 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 1/1000 - 1/10000. Detects a band of approximately 68 kDa (predicted molecular weight: 52 kDa).
IHC-P 1/50 - 1/100.

Perform heat mediated antigen retrieval using Citrate buffer, pH 6.0.

ICC/IF 1/500.

For unpurified format use at 1/50 - 1/100 dilution.

IP 1/20.
Flow Cyt 1/30.
IHC-P 1/50. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

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.
  • Target information above from: UniProt accession Q13283 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 10146 Human
    • Entrez Gene: 27041 Mouse
    • Entrez Gene: 171092 Rat
    • Omim: 608431 Human
    • SwissProt: Q13283 Human
    • SwissProt: P97855 Mouse
    • Unigene: 587054 Human
    • Unigene: 380129 Mouse
    • Unigene: 39631 Mouse
    • Unigene: 405739 Mouse
    • Unigene: 25201 Rat
    see all
  • 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 [EPR13986(B)] (ab181150)
    Western blot - Anti-G3BP antibody [EPR13986(B)] (ab181150)
    All lanes : Anti-G3BP antibody [EPR13986(B)] (ab181150) at 1/1000 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 - ab181150 observed at 68 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab181150 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. Ab181150 and ab8245 (Mouse anti-GAPDH 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 1/20000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-G3BP antibody [EPR13986(B)] (ab181150)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-G3BP antibody [EPR13986(B)] (ab181150)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human hepatocellular cancer tissue sections labeling G3BP with purified ab181150 at 1/50 dilution (6.86 µg/ml). Heat mediated antigen retrieval was performed using Citrate buffer, pH 6.0. ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

  • Western blot - Anti-G3BP antibody [EPR13986(B)] (ab181150)
    Western blot - Anti-G3BP antibody [EPR13986(B)] (ab181150)
    All lanes : Anti-G3BP antibody [EPR13986(B)] (ab181150) at 1/10000 dilution (Purified)

    Lane 1 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates
    Lane 2 : Ramos (Human Burkitt's lymphoma B lymphocyte) whole cell lysates
    Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates
    Lane 4 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

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

  • Immunocytochemistry/ Immunofluorescence - Anti-G3BP antibody [EPR13986(B)] (ab181150)
    Immunocytochemistry/ Immunofluorescence - Anti-G3BP antibody [EPR13986(B)] (ab181150)

    Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling G3BP with purified ab181150 at 1/500 dilution (0.7 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • Western blot - Anti-G3BP antibody [EPR13986(B)] (ab181150)
    Western blot - Anti-G3BP antibody [EPR13986(B)] (ab181150)
    All lanes : Anti-G3BP antibody [EPR13986(B)] (ab181150) at 1/5000 dilution (unpurified)

    Lane 1 : Jurkat cell lysate with NFDM/TBST
    Lane 2 : Ramos 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?

  • Immunoprecipitation - Anti-G3BP antibody [EPR13986(B)] (ab181150)
    Immunoprecipitation - Anti-G3BP antibody [EPR13986(B)] (ab181150)

    ab181150 (purified) at 1/20 dilution (2ug) immunoprecipitating G3BP in Ramos whole cell lysate. Ramos (Human Burkitt's lymphoma B lymphocyte) whole cell lysate 10ug
    Lane 2 (+): ab181150 & Ramos whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab181150 in Ramos whole cell lysate
    For western blotting, VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/1000 dilution.
    Blocking and diluting buffer: 5% NFDM/TBST.

  • Flow Cytometry - Anti-G3BP antibody [EPR13986(B)] (ab181150)
    Flow Cytometry - Anti-G3BP antibody [EPR13986(B)] (ab181150)

    Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling G3BP with purified ab181150 at 1/30 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-G3BP antibody [EPR13986(B)] (ab181150)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-G3BP antibody [EPR13986(B)] (ab181150)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat kidney tissue sections labeling G3BP with purified ab181150 at 1/50 dilution (6.86 µg/ml). Heat mediated antigen retrieval was performed using Citrate buffer, pH 6.0. ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-G3BP antibody [EPR13986(B)] (ab181150)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-G3BP antibody [EPR13986(B)] (ab181150)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse kidney tissue sections labeling G3BP with purified ab181150 at 1/50 dilution (6.86 µg/ml). Heat mediated antigen retrieval was performed using Citrate buffer, pH 6.0. ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

  • Immunocytochemistry/ Immunofluorescence - Anti-G3BP antibody [EPR13986(B)] (ab181150)
    Immunocytochemistry/ Immunofluorescence - Anti-G3BP antibody [EPR13986(B)] (ab181150)

    Immunofluorescent analysis of 4% paraformaldehyde fixed 293 cells staining G3BP using ab181150 (unpurified) at 1/100 dilution, and Alexa Fluor®555 stained Goat anti Rabbit IgG at 1/200 dilution as a secondary antibody (red). Dapi counterstain (blue)

  • Western blot - Anti-G3BP antibody [EPR13986(B)] (ab181150)
    Western blot - Anti-G3BP antibody [EPR13986(B)] (ab181150)
    All lanes : Anti-G3BP antibody [EPR13986(B)] (ab181150) at 1/1000 dilution (unpurified)

    Lane 1 : Hela cell lysate with NFDM/TBST
    Lane 2 : 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 [EPR13986(B)] (ab181150)
    Immunohistochemistry paraffin embedded sections - Anti-G3BP antibody [EPR13986(B)] (ab181150)

    Immunohistochemical analysis of paraffin-embedded Human colon tissue staining G3BP using ab181150 (unpurified) at 1/100 dilution, and prediluted HRP Polymer for Rabbit IgG as a secondary antibody with Hematoxylin counterstain.

  • Immunoprecipitation - Anti-G3BP antibody [EPR13986(B)] (ab181150)
    Immunoprecipitation - Anti-G3BP antibody [EPR13986(B)] (ab181150)

    Western blot analysis of G3BP in immunoprecipitation pellets from Ramos cell lysate, using ab181151 at a 1/50 dilution. Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated was used as a secondary antibody at 1/1000 dilution. Blocking/dilution buffer and concentration: 5% NFDM/TBST



    Anti-G3BP antibody [EPR13986(B)] (ab181150) at 1/50 dilution (unpurified) + Immunoprecipitate from Ramos cell lysate using ab181150 with NFDM/TBST at 5 %

    Secondary
    Peroxidase conjugated Goat anti-Rabbit IgG (H+L) at 1/1000 dilution
  • Anti-G3BP antibody [EPR13986(B)] (ab181150)
    Anti-G3BP antibody [EPR13986(B)] (ab181150)

Protocols

  • Immunoprecipitation protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

    • Datasheet

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

  • References (8)

    Publishing research using ab181150? Please let us know so that we can cite the reference in this datasheet.

    ab181150 has been referenced in 8 publications.

    • Pla-Martín D  et al. CLUH granules coordinate translation of mitochondrial proteins with mTORC1 signaling and mitophagy. EMBO J 39:e102731 (2020). PubMed: 32149416
    • Cui X  et al. Homer1 is a Potential Biomarker for Prognosis in Human Colorectal Carcinoma, Possibly in Association with G3BP1 Signaling. Cancer Manag Res 12:2899-2909 (2020). PubMed: 32425603
    • Das R  et al. New roles for the de-ubiquitylating enzyme OTUD4 in an RNA-protein network and RNA granules. J Cell Sci 132:N/A (2019). PubMed: 31138677
    • Herman AB  et al. Regulation of Stress Granule Formation by Inflammation, Vascular Injury, and Atherosclerosis. Arterioscler Thromb Vasc Biol 39:2014-2027 (2019). PubMed: 31462091
    • Berges N  et al. Complex alternative splicing of human Endonuclease V mRNA, but evidence for only a single protein isoform. PLoS One 14:e0225081 (2019). PubMed: 31703097
    • Hartmann H  et al. Proteomics and C9orf72 neuropathology identify ribosomes as poly-GR/PR interactors driving toxicity. Life Sci Alliance 1:e201800070 (2018). PubMed: 30456350
    • Zhang Y  et al. Downregulated miR-621 promotes cell proliferation via targeting CAPRIN1 in hepatocellular carcinoma. Am J Cancer Res 8:2116-2129 (2018). PubMed: 30416861
    • 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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