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Cell Biology Proteolysis / Ubiquitin Proteasome / Ubiquitin Ubiquitin E3 Enzymes RING Finger E3 Ligase
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-TRIM25/EFP antibody [EPR7315] (ab167154)

  • Datasheet
Reviews (1) Submit a question References (13)

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Western blot - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)
  • Western blot - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)
  • Immunocytochemistry/ Immunofluorescence - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)
  • Flow Cytometry - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)
  • Western blot - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)
  • Immunoprecipitation - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)
  • Western blot - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)
  • Western blot - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)
  • Immunocytochemistry/ Immunofluorescence - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)
  • Anti-TRIM25/EFP antibody [EPR7315] (ab167154)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR7315] to TRIM25/EFP
  • Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

You may also be interested in

Preparation
Product image
Mitochondria Isolation Kit for Cultured Cells (ab110170)
Protein
Product image
Recombinant Human TRIM25/EFP protein (ab152814)
Secondary
Product image
Goat Anti-Rabbit IgG H&L (HRP) (ab205718)

View more associated products

Overview

  • Product name

    Anti-TRIM25/EFP antibody [EPR7315]
    See all TRIM25/EFP primary antibodies
  • Description

    Rabbit monoclonal [EPR7315] to TRIM25/EFP
  • 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/EFP aa 100-200. The exact sequence is proprietary.
    Database link: Q14258

  • Positive control

    • Recombinant Human TRIM25/EFP 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

     This product was previously labelled as TRIM25

     

    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. Stable for 12 months at -20°C.
  • Storage buffer

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

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR7315
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Proteolysis / Ubiquitin
    • Proteasome / Ubiquitin
    • Ubiquitin E3 Enzymes
    • RING Finger E3 Ligase
    • Signal Transduction
    • Growth Factors/Hormones
    • Hormones
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Co-factors

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-TRIM25/EFP antibody [EPR7315] (ab200781)
    • HRP Anti-TRIM25/EFP antibody [EPR7315] (ab200788)
    • Anti-TRIM25/EFP antibody [EPR7315] - BSA and Azide free (ab232356)
  • 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)
    • Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

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

    • HeLa whole cell lysate (ab150035)
    • Recombinant Human TRIM25/EFP protein (ab152814)
    • HeLa whole cell lysate (ab29545)
    • K562 whole cell lysate (ab7911)
  • Recombinant Protein

    • Recombinant Human TRIM25/EFP protein (ab152814)

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. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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.
  • Target information above from: UniProt accession Q14258 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 7706 Human
    • Entrez Gene: 217069 Mouse
    • Entrez Gene: 494338 Rat
    • Omim: 600453 Human
    • SwissProt: Q14258 Human
    • SwissProt: Q61510 Mouse
    • Unigene: 528952 Human
    • Unigene: 248445 Mouse
    • Unigene: 34382 Rat
    see all
  • 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

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

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : TRIM25/EFP 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/EFP in wild-type HAP1 cells as signal was lost in TRIM25/EFP knockout cells. Wild-type and TRIM25/EFP 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.

  • Western blot - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)
    Western blot - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)
    All lanes : Anti-TRIM25/EFP 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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)

    ab167154 staining TRIM25/EFP 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.

     

  • Immunocytochemistry/ Immunofluorescence - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)
    Immunocytochemistry/ Immunofluorescence - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)

    ab167154 staining TRIM25/EFP 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 - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)
    Flow Cytometry - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)

    Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling TRIM25/EFP 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.

  • Western blot - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)
    Western blot - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)
    Anti-TRIM25/EFP 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 

  • Immunoprecipitation - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)
    Immunoprecipitation - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)

    ab167154 immunoprecipitating TRIM25/EFP. 10µg of cell lysate was incubated with primary antibody at a dilution of 1/40 and VeriBlot for IP Detection Reagent (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.

  • Western blot - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)
    Western blot - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)
    Anti-TRIM25/EFP 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

  • Western blot - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)
    Western blot - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)
    All lanes : Anti-TRIM25/EFP 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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)

    Immunohistochemical analysis of paraffin-embedded Human breast tissue labeling TRIM25/EFP with ab167154 at 1/100 dilution.

  • Immunocytochemistry/ Immunofluorescence - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)
    Immunocytochemistry/ Immunofluorescence - Anti-TRIM25/EFP antibody [EPR7315] (ab167154)

    Immunofluorescent analysis of HeLa cells labeling TRIM25/EFP with ab167154 at 1/100 dilution.

  • Anti-TRIM25/EFP antibody [EPR7315] (ab167154)
    Anti-TRIM25/EFP antibody [EPR7315] (ab167154)

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
  • References (13)

    Publishing research using ab167154? Please let us know so that we can cite the reference in this datasheet.

    ab167154 has been referenced in 13 publications.

    • Gupta S  et al. Interaction With 14-3-3 Correlates With Inactivation of the RIG-I Signalosome by Herpesvirus Ubiquitin Deconjugases. Front Immunol 11:437 (2020). PubMed: 32226432
    • Marcos-Villar L  et al. Interferon-? Stimulation Elicited by the Influenza Virus Is Regulated by the Histone Methylase Dot1L through the RIG-I-TRIM25 Signaling Axis. Cells 9:N/A (2020). PubMed: 32188146
    • Garcia-Moreno M  et al. System-wide Profiling of RNA-Binding Proteins Uncovers Key Regulators of Virus Infection. Mol Cell 74:196-211.e11 (2019). PubMed: 30799147
    • Ficarelli M  et al. KHNYN is essential for the zinc finger antiviral protein (ZAP) to restrict HIV-1 containing clustered CpG dinucleotides. Elife 8:N/A (2019). PubMed: 31284899
    • Gupta S  et al. 14-3-3 scaffold proteins mediate the inactivation of trim25 and inhibition of the type I interferon response by herpesvirus deconjugases. PLoS Pathog 15:e1008146 (2019). PubMed: 31710640
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    Western blot abreview for Anti-TRIM25/EFP antibody [EPR7315]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Dopaminergic Neurons)
    Gel Running Conditions
    Reduced Denaturing (NuPage 4-12% MES buffer)
    Loading amount
    30 µg
    Specification
    Dopaminergic Neurons
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 20 2020

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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