Recombinant Anti-TRIM25/EFP antibody [EPR7315] - BSA and Azide free (ab232356)
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 - BSA and Azide free
- Suitable for: Flow Cyt (Intra), ICC/IF, IP, IHC-P, WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-TRIM25/EFP antibody [EPR7315] - BSA and Azide free
See all TRIM25/EFP primary antibodies -
Description
Rabbit monoclonal [EPR7315] to TRIM25/EFP - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), ICC/IF, IP, IHC-P, WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HAP1, HeLa, and MCF7 whole cell lysates. IHC-P: Human pancreas tissue.
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General notes
ab232356 is the carrier-free version of ab167154.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR7315 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab232356 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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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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ICC/IF |
Use at an assay dependent concentration.
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IP |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 71 kDa.
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Notes |
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Flow Cyt (Intra)
Use at an assay dependent concentration. |
ICC/IF
Use at an assay dependent concentration. |
IP
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 71 kDa. |
Target
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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
modificationsAuto-ISGylated. -
Cellular localization
Cytoplasm. Colocalized with DDX58 at cytoplasmic perinuclear bodies. - Information by UniProt
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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
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Alternative names
- E3 ubiquitin/ISG15 ligase TRIM25 antibody
- EFP antibody
- Estrogen responsive finger protein antibody
see all
Images
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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 kDaThis data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab167154).
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. -
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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab167154).
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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.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab167154).
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Immunocytochemistry/ Immunofluorescence - Anti-TRIM25/EFP antibody [EPR7315] - BSA and Azide free (ab232356)
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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab167154).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRIM25/EFP antibody [EPR7315] - BSA and Azide free (ab232356)
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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab167154).
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (0)
ab232356 has not yet been referenced specifically in any publications.