Anti-G3BP antibody (ab56574)

Mouse monoclonal G3BP antibody. Validated in WB, IHC, Flow Cyt, ICC/IF and tested in Human. Cited in 13 publication(s). Independently reviewed in 7 review(s).

Overview

  • Product name

  • Description

    Mouse monoclonal to G3BP
  • Host species

    Mouse
  • Tested applications

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

    Reacts with: Human
  • Immunogen

    Recombinant fragment: KPEPVLEETA PEDAQKSSSP APADIAQTVQ EDLRTFSWAS VTSKNLPPSG AVPVTGIPPH VVKVPASQPR PESKPESQIP PQRPQRDQRV , corresponding to amino acids 214-303 of Human G3BP

  • General notes

    This product was changed from ascites to tissue culture supernatant on 22/03/2019. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

Properties

Applications

Our Abpromise guarantee covers the use of ab56574 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 Use at an assay dependent concentration. Predicted molecular weight: 52 kDa.
IHC-P Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

ICC/IF Use at an assay dependent concentration.

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

  • ICC/IF image of ab56574 stained HeLa cells. The cells were 4% formaldehyde 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 (ab56574, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 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 image was generated using the ascites version of the product.

  • G3BP antibody (ab56574) used in immunohistochemistry at 1ug/ml on formalin fixed and paraffin embedded human lymphoma.

    This image was generated using the ascites version of the product.

  • G3BP antibody (ab56574) at 1ug/lane + A-431 cell lysate at 25ug/lane.

    This image was generated using the ascites version of the product.

  • Overlay histogram showing HeLa cells stained with ab56574 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab56574, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 80% methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

    This image was generated using the ascites version of the product.

  • ab56574 staining G3BP in Human HeLa cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with Triton X-100 and blocked with 5% BSA for 12 hours at 4°C. Samples were incubated with primary antibody (1/500 in PBS) for 1 hour at 37°C. An Alexa Fluor® 488-conjugated Goat anti-mouse IgG polyclonal (1/1000) was used as the secondary antibody.

    Top row - untreated cells. Bottom row - cells treated with sodium arsenite. Left - G3BP, Middle - Nucleus, Right - Merge.

    Stress granules are visible in cells treated with sodium arsenite, whereas G3BP is dispersed in the cytoplasm in untreated cells.

    This image was generated using the ascites version of the product.

    See Abreview

References

This product has been referenced in:

  • Tauber D & Parker R 15-Deoxy-?12,14-prostaglandin J2 promotes phosphorylation of eukaryotic initiation factor 2a and activates the integrated stress response. J Biol Chem 294:6344-6352 (2019). Read more (PubMed: 30723157) »
  • Christen KE  et al. Psammaplysin F increases the efficacy of bortezomib and sorafenib through regulation of stress granule formation. Int J Biochem Cell Biol 112:24-38 (2019). Read more (PubMed: 31022461) »
See all 23 Publications for this product

Customer reviews and Q&As

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1-7 of 7 Abreviews

Application
Western blot
Sample
Human Cell lysate - whole cell (Cancer Cell lines)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
20 µg
Specification
Cancer Cell lines
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Dec 13 2017

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Epithelial cells)
Permeabilization
Yes - Triton-X 100
Specification
Epithelial cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 20°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Apr 07 2017

Application
Western blot
Sample
Human Cell lysate - whole cell (Epithelial cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 µg
Specification
Epithelial cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Mar 27 2017

Application
Western blot
Loading amount
60 µg
Gel Running Conditions
Reduced Denaturing
Sample
Human Cell lysate - whole cell (epithelial)
Specification
epithelial
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Sep 16 2014

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 4°C
Sample
Human Cell (epithelial cells)
Specification
epithelial cells
Permeabilization
Yes - Triton x100
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Aug 25 2014

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Sample
Human Cell (Hela Cells)
Specification
Hela Cells
Permeabilization
Yes - Triton x100
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Aug 23 2013

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Hepatocytes)
Specification
Hepatocytes
Fixative
Paraformaldehyde
Permeabilization
No
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Oct 04 2012

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