Name: Anti-TRIM25/EFP antibody [EPR7315]
Brand: RabMAb
SKU: ab167154
Rating: 5 (1 reviews)

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Western blot - 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: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP
Knockout validated
Reacts with: Mouse, Rat, Human

Alexa Fluor® 488 Carrier Free HRP

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: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IPmore 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 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.

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

Alternative Versions
Compatible Secondaries
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
Recombinant Protein
- Recombinant Human TRIM25/EFP protein (ab152814)

The Abpromise guarantee

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
Flow Cyt (Intra)		1/60 - 1/90.
WB	(1)	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.

Notes
Flow Cyt (Intra) 1/60 - 1/90.
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.

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
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

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)

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)

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)

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

Intracellular 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)

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)

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)

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)

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)

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)

Immunofluorescent analysis of HeLa cells labeling TRIM25/EFP with ab167154 at 1/100 dilution.
Anti-TRIM25/EFP antibody [EPR7315] (ab167154)

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

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

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

ab167154 has been referenced in 26 publications.

D'Amico G et al. ERG activity is regulated by endothelial FAK coupling with TRIM25/USP9x in vascular patterning. Development 149:N/A (2022). PubMed: 35723257
Booy EP et al. The noncoding RNA BC200 associates with polysomes to positively regulate mRNA translation in tumor cells. J Biol Chem 296:100036 (2021). PubMed: 33410401
Mei P et al. E3 ligase TRIM25 ubiquitinates RIP3 to inhibit TNF induced cell necrosis. Cell Death Differ 28:2888-2899 (2021). PubMed: 33953350
Kumar A et al. SARS-CoV-2 Nonstructural Protein 1 Inhibits the Interferon Response by Causing Depletion of Key Host Signaling Factors. J Virol 95:e0026621 (2021). PubMed: 34110264
Smith EM et al. The mTOR regulated RNA-binding protein LARP1 requires PABPC1 for guided mRNA interaction. Nucleic Acids Res 49:458-478 (2021). PubMed: 33332560

View all Publications for this product

Customer reviews and Q&As

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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

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Submitted Oct 20 2020

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Conjugation kits

Isotype control

Positive Controls

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Pathway

Sequence similarities

Domain

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

References (26)

Customer reviews and Q&As

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

Post-translational
modifications