Recombinant 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, ICC/IF, IP, Flow Cyt, IHC-P
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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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, ICC/IF, IP, Flow Cyt, IHC-Pmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Wild-type A431, Jurkat and HEK-293 whole cell lysates.
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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
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
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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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR13985(B) -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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KO cell lysates
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Positive Controls
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Recombinant Protein
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Related Products
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 |
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WB |
1/10000 - 1/50000. Detects a band of approximately 68 kDa (predicted molecular weight: 52 kDa).
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ICC/IF |
1/250 - 1/500.
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IP |
1/20 - 1/40.
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Flow Cyt |
1/20.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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IHC-P |
1/50 - 1/100.
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Notes |
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WB
1/10000 - 1/50000. Detects a band of approximately 68 kDa (predicted molecular weight: 52 kDa). |
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. |
IHC-P
1/50 - 1/100. |
Target
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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
modificationsPhosphorylated 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
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Database links
- Entrez Gene: 10146 Human
- Omim: 608431 Human
- SwissProt: Q13283 Human
- Unigene: 587054 Human
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Alternative names
- ATP dependent DNA helicase VIII antibody
- ATP-dependent DNA helicase VIII antibody
- G3BP antibody
see all
Images
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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) 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.
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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.
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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 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).
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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.
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All lanes : Anti-G3BP antibody [EPR13985(B)] (ab181149) at 1/10000 dilution
Lane 1 : Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 2 : G3BP1 knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 3 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 4 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 52 kDaLanes 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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (8)
ab181149 has been referenced in 8 publications.
- Da Ros M et al. Wild-type and mutant SOD1 localizes to RNA-rich structures in cells and mice but does not bind RNA. J Neurochem 156:524-538 (2021). PubMed: 32683701
- 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