Recombinant Anti-TRIM56 antibody [EPR10583] (ab154862)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR10583] to TRIM56
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-TRIM56 antibody [EPR10583]
See all TRIM56 primary antibodies -
Description
Rabbit monoclonal [EPR10583] to TRIM56 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human TRIM56. The exact sequence is proprietary.
Database link: Q9BRZ2 -
Positive control
- IHC-P: Human brain, pancreas and colon cancer tissues. WB: HAP1, A549, A375, MCF7 and HeLa cell lysates. ICC/IF: MCF7 and HeLa cells Flow Cyt (intra): MCF7 cells
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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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR10583 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab154862 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) |
1/20.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. For unpurified use at 1/100 - 1/500 dilution. |
|
WB |
1/10000 - 1/50000. Predicted molecular weight: 81 kDa.
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IHC-P |
1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. For unpurifed use at 1/50 -1/100 dilution. |
|
ICC/IF |
1/70.
For unpurified use at 1/100 - 1/250 dilution. |
Notes |
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Flow Cyt (Intra)
1/20. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. For unpurified use at 1/100 - 1/500 dilution. |
WB
1/10000 - 1/50000. Predicted molecular weight: 81 kDa. |
IHC-P
1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. See IHC antigen retrieval protocols. For unpurifed use at 1/50 -1/100 dilution. |
ICC/IF
1/70. For unpurified use at 1/100 - 1/250 dilution. |
Target
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Function
E3 ubiquitin-protein ligase that mediates 'Lys-63'-linked polyubiquitination of TMEM173/STING, thereby playing a key role in innate immunity. TMEM173/STING 'Lys-63'-linked ubiquitination activates the production of type I interferon IFN-beta following detection of pathogen- and host-derived double-stranded DNA. -
Sequence similarities
Belongs to the TRIM/RBCC family.
Contains 2 B box-type zinc fingers.
Contains 1 RING-type zinc finger. -
Cellular localization
Cytoplasm. - Information by UniProt
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Database links
- Entrez Gene: 81844 Human
- SwissProt: Q9BRZ2 Human
- Unigene: 521092 Human
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Alternative names
- A130009K11Rik antibody
- DKFZp667O116 antibody
- E3 ubiquitin-protein ligase TRIM56 antibody
see all
Images
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All lanes : Anti-TRIM56 antibody [EPR10583] (ab154862) at 1/1000 dilution
Lane 1 : Wild-type A549 (Human lung carcinoma cell line) whole cell lysate
Lane 2 : TRIM56 knockout A549 (Human lung carcinoma cell line) whole cell lysate
Lane 3 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
Lane 4 : A-375 (Human malignant melanoma cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 81 kDa
Observed band size: 88 kDa why is the actual band size different from the predicted?Lanes 1-4: Merged signal (red and green). Green - ab154862 observed at 88 kDa. Red - loading control ab8245 observed at 36 kDa.
ab154862 Anti-TRIM56 antibody [EPR10583] was shown to specifically react with TRIM56 in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267063 (knockout cell lysate ab258250) was used. Wild-type and TRIM56 knockout samples were subjected to SDS-PAGE. ab154862 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling TRIM56 with purified ab154862 at 1/70 dilution (10 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 (2.5 µg/ml) dilution. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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Intracellular Flow Cytometry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling TRIM56 with purified ab154862 at 1/70 dilution (10µg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colon cancer tissue sections labeling TRIM56 with purified ab154862 at 1/1000 dilution (0.691 µg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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All lanes : Anti-TRIM56 antibody [EPR10583] (ab154862) at 1/1000 dilution
Lane 1 : Wild-type A549 (Human lung carcinoma cell line) whole cell lysate
Lane 2 : TRIM56 knockout A549 (Human lung carcinoma cell line) whole cell lysate
Lane 3 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
Lane 4 : A-375 (Human malignant melanoma cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 81 kDa
Observed band size: 88 kDa why is the actual band size different from the predicted?Lanes 1-4: Merged signal (red and green). Green - ab154862 observed at 88 kDa. Red - loading control ab8245 observed at 36 kDa.
ab154862 Anti-TRIM56 antibody [EPR10583] was shown to specifically react with TRIM56 in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267062 (knockout cell lysate ab258249) was used. Wild-type and TRIM56 knockout samples were subjected to SDS-PAGE. ab154862 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-TRIM56 antibody [EPR10583] (ab154862) at 1/50000 dilution (Purified)
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
Lane 2 : A375 (Human malignant melanoma epithelial cell) whole cell lysates
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 81 kDa
Observed band size: 81 kDa -
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: TRIM56 knockout HAP1 cell lysate (20 µg)
Lane 3: MCF7 cell lysate (20 µg)
Lane 4: A375 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab154862 observed at 88 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab154862 (unpurifed) was shown to recognize TRIM56 when TRIM56 knockout samples were used, along with additional cross-reactive bands. Wild-type and TRIM56 knockout samples were subjected to SDS-PAGE. ab154862 and ab8245 (loading control to GAPDH) were both diluted 1/10000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging. -
All lanes : Anti-TRIM56 antibody [EPR10583] (ab154862) at 1/10000 dilution ((unpurified))
Lane 1 : HeLa cell lysate
Lane 2 : A375 cell lysate
Lane 3 : MCF7 cell lysate
Lysates/proteins at 10 µg per lane.
Predicted band size: 81 kDa -
Immunohistochemical analysis of paraffin-embedded Human brain tissue labeling TRIM56 with ab154862 (unpurified) at 1/50.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling TRIM56 with ab154862 (unpurified) at 1/50.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunofluorescent analysis of MCF7 cells labeling TRIM56 with ab154862 (unpurified) at 1/100.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (6)
ab154862 has been referenced in 6 publications.
- Yang X et al. TRIM56 promotes malignant progression of glioblastoma by stabilizing cIAP1 protein. J Exp Clin Cancer Res 41:336 (2022). PubMed: 36471347
- Yang P et al. G3BP1 Is a Tunable Switch that Triggers Phase Separation to Assemble Stress Granules. Cell 181:325-345.e28 (2020). PubMed: 32302571
- 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
- Xue M et al. Regulation of estrogen signaling and breast cancer proliferation by an ubiquitin ligase TRIM56. Oncogenesis 8:30 (2019). PubMed: 31000690
- Sikorski K et al. A high-throughput pipeline for validation of antibodies. Nat Methods 15:909-912 (2018). PubMed: 30377371
- Fiskin E et al. Structural basis for the recognition and degradation of host TRIM proteins by Salmonella effector SopA. Nat Commun 8:14004 (2017). WB . PubMed: 28084320