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  1. Link

    g3bp-antibody-epr13985b-ab181149.pdf

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Epigenetics and Nuclear Signaling DNA / RNA DNA / Nucleotides
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Validated using a knockout cell lineRecombinantRabMAb

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

  • Datasheet
  • Certificate of Compliance
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Western blot - Anti-G3BP antibody [EPR13985(B)] (ab181149)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-G3BP antibody [EPR13985(B)] (ab181149)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-G3BP antibody [EPR13985(B)] (ab181149)
  • Immunoprecipitation - Anti-G3BP antibody [EPR13985(B)] (ab181149)
  • Flow Cytometry - Anti-G3BP antibody [EPR13985(B)] (ab181149)
  • Immunocytochemistry - Anti-G3BP antibody [EPR13985(B)] (ab181149)
  • Western blot - 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, IP, Flow Cyt
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free

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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, IP, Flow Cytmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

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

  • Positive control

    • WB: Wild-type A431, Jurkat and HEK-293 whole cell lysates.
  • 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.

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 (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR13985(B)
  • Isotype

    IgG
  • Research areas

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

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-G3BP antibody [EPR13985(B)] (ab216928)
    • Alexa Fluor® 647 Anti-G3BP antibody [EPR13985(B)] (ab217087)
    • Anti-G3BP antibody [EPR13985(B)] - BSA and Azide free (ab240246)
  • Compatible Secondaries

    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human G3BP1 knockout A-431 cell line (ab261886)
  • KO cell lysates

    • Human G3BP1 knockout A-431 cell lysate (ab261695)
  • 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

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.
IP
1/20 - 1/40.
Flow Cyt
1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

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.
IP
1/20 - 1/40.
Flow Cyt
1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

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
    • Omim: 608431 Human
    • SwissProt: Q13283 Human
    • Unigene: 587054 Human
    • 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 1/20000 dilution (Purified)

      Lane 1 : Ramos (Human Burkitt's lymphoma B lymphocyte) whole cell lysate
      Lane 2 : Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysate

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

      Predicted band size: 52 kDa

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

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue sections labeling G3BP with Purified ab181149 at 1:300 (0.4 µg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control. 

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

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast tissue sections labeling G3BP with Purified ab181149 at 1:300 (0.4 µg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control. 

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

      Purified ab181149 at 1:20 dilution (0.6µg) immunoprecipitating G3BP in Ramos whole cell lysate.
      Lane 1 (input): Ramos (Human Burkitt's lymphoma B lymphocyte) whole cell lysate 10 µg.
      Lane 2 (+): ab181149 + Ramos whole cell lysate.
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab181149 in Ramos whole cell lysate.
      VeriBlot for IP Detection Reagent (HRP)(ab131366) (1:1000 dilution) was used for Western blotting.
      Blocking Buffer and concentration: 5% NFDM/TBST.
      Diluting buffer and concentration: 5% NFDM/TBST.
      Observed band size: kDa

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

      Flow Cytometry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labelling G3BP with Purified ab181149 at 1:20 dilution (5 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150081) secondary antibody was used at 1:2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).

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

      Immunocytochemistry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling G3BP with Purified ab181149 at 1:50 dilution (2.4 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]+H21:L21 - Microtubule Marker (Alexa Fluor® 594) 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 [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.

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

    Protocols

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

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    Certificate of Compliance

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

    References (7)

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

    ab181149 has been referenced in 7 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
    • Lee AK  et al. Translational Repression of G3BP in Cancer and Germ Cells Suppresses Stress Granules and Enhances Stress Tolerance. Mol Cell 79:645-659.e9 (2020). PubMed: 32692974
    • 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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