Overview

  • Product name
    Anti-TRIM25 antibody [EPR7315]
    See all TRIM25 primary antibodies
  • Description
    Rabbit monoclonal [EPR7315] to TRIM25
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human TRIM25. The exact sequence is proprietary.

  • Positive control
    • Recombinant Human TRIM25 protein (ab152814) can be used as a positive control in WB. HeLa and K562 cell lysates; Human breast tissue. Human pancreas tissue. Rat brain lysate. SP2/0 (mouse spleen) whole cell lysate.
  • General notes

    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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab167154 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 1/2000 - 1/50000. Predicted molecular weight: 71 kDa.
IHC-P 1/100 - 1/250.
ICC/IF 1/500 - 1/1000.
IP 1/40 - 1/100.
Flow Cyt 1/60 - 1/90.

Target

  • Function
    Functions as an ubiquitin E3 ligase and as an ISG15 E3 ligase. Involved in innate immune defense against viruses by mediating ubiquitination of DDX58. Mediates 'Lys-63'-linked polyubiquitination of the DDX58 N-terminal CARD-like region which is crucial for triggering the cytosolic signal transduction that leads to the production of interferons in response to viral infection. Promotes ISGylation of 14-3-3 sigma (SFN), an adapter protein implicated in the regulation of a large spectrum signaling pathway. Mediates estrogen action in various target organs.
  • Tissue specificity
    Ubiquitous.
  • Pathway
    Protein modification; protein ubiquitination.
  • Sequence similarities
    Contains 1 B30.2/SPRY domain.
    Contains 1 RING-type zinc finger.
  • Domain
    The RING-type zinc finger is important for ISG15 E3 ligase activity and autoISGylation. AutoISGylation negatively regulates ISG15 E3 ligase activity.
    The C-terminal B30.2/SPRY domain interacts with the first N-terminal CARD domain of DDX58.
  • Post-translational
    modifications
    Auto-ISGylated.
  • Cellular localization
    Cytoplasm. Colocalized with DDX58 at cytoplasmic perinuclear bodies.
  • Information by UniProt
  • Database links
  • Alternative names
    • E3 ubiquitin/ISG15 ligase TRIM25 antibody
    • EFP antibody
    • Estrogen responsive finger protein antibody
    • Estrogen-responsive finger protein antibody
    • RING finger protein 147 antibody
    • RNF 147 antibody
    • RNF147 antibody
    • TRI25 antibody
    • TRI25_HUMAN antibody
    • TRIM 25 antibody
    • Trim25 antibody
    • Tripartite motif containing 25 antibody
    • Tripartite motif containing protein 25 antibody
    • Tripartite motif protein TRIM25 antibody
    • Tripartite motif-containing protein 25 antibody
    • Tripartite motifcontaining25 antibody
    • Ubiquitin/ISG15-conjugating enzyme TRIM25 antibody
    • Z147 antibody
    • Zinc finger protein 147 (estrogen responsive finger protein) antibody
    • Zinc finger protein 147 antibody
    • Zinc finger protein-147 antibody
    • ZNF 147 antibody
    • ZNF147 antibody
    see all

Images

  • All lanes : Anti-TRIM25 antibody [EPR7315] (ab167154) at 1/10000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : TRIM25 knockout HAP1 whole cell lysate
    Lane 3 : HeLa whole cell lysate
    Lane 4 : MCF7 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 71 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab167154 observed at 71 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab167154 was shown to specifically react with TRIM25 in wild-type HAP1 cells as signal was lost in TRIM25 knockout cells. Wild-type and TRIM25 knockout samples were subjected to SDS-PAGE. Ab167154 and ab9484 (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.

  • All lanes : Anti-TRIM25 antibody [EPR7315] (ab167154) at 1/50000 dilution

    Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate

    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: 71 kDa



    Blocking and diluting buffer: 5% NFDM/TBST

  • ab167154 staining TRIM25 in human pancreas tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

     

  • ab167154 staining TRIM25 in HeLa (human cervix adenocarcinoma) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/500. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a dilution of 1/1000. ab195889 was used as a tubulin counterstain at a dilution of 1/200 and DAPI was used as a nuclear counterstain.

  • Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling TRIM25 with purified ab167154 at 1/90 dilution(10ug/mL) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

  • Anti-TRIM25 antibody [EPR7315] (ab167154) at 1/2000 dilution + SP2/0 (mouse spleen) whole cell lysate at 15 µg

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

    Predicted band size: 71 kDa
    Additional bands at: 71 kDa. We are unsure as to the identity of these extra bands.



    Blocking and diluting buffer: 5% NFDM /TBST 

  • ab167154 immunoprecipitating TRIM25. 10µg of cell lysate was incubated with primary antibody at a dilution of 1/40 and VeriBlot for IP secondary antibody (HRP) (ab131366) at a dilution of 1/1000.

    Lane 1: HeLa (human cervix adenocarcinoma) whole cell lysate (10ug)
    Lane 2: HeLa (human cervix adenocarcinoma) whole cell lysate
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab167154 in HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate.

  • Anti-TRIM25 antibody [EPR7315] (ab167154) at 1/10000 dilution + Rat brain lysate at 15 µg

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

    Predicted band size: 71 kDa
    Additional bands at: 71 kDa. We are unsure as to the identity of these extra bands.



    Blocking and diluting buffer: 5% NFDM/TBST

  • All lanes : Anti-TRIM25 antibody [EPR7315] (ab167154) at 1/10000 dilution

    Lane 1 : HeLa cell lysate
    Lane 2 : K562 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP labeled goat anti-rabbit at 1/2000 dilution

    Predicted band size: 71 kDa

  • Immunohistochemical analysis of paraffin-embedded Human breast tissue labeling TRIM25 with ab167154 at 1/100 dilution.
  • Immunofluorescent analysis of HeLa cells labeling TRIM25 with ab167154 at 1/100 dilution.

References

This product has been referenced in:
  • Gupta S  et al. Herpesvirus deconjugases inhibit the IFN response by promoting TRIM25 autoubiquitination and functional inactivation of the RIG-I signalosome. PLoS Pathog 14:e1006852 (2018). Read more (PubMed: 29357390) »
  • Choudhury NR  et al. RNA-binding activity of TRIM25 is mediated by its PRY/SPRY domain and is required for ubiquitination. BMC Biol 15:105 (2017). WB . Read more (PubMed: 29117863) »
See all 6 Publications for this product

Customer reviews and Q&As

There are currently no Customer reviews or Questions for ab167154.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Sign up