Recombinant Anti-TRIM21/SS-A antibody [EPR20290] (ab207728)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20290] to TRIM21/SS-A
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-TRIM21/SS-A antibody [EPR20290]
See all TRIM21/SS-A primary antibodies -
Description
Rabbit monoclonal [EPR20290] to TRIM21/SS-A -
Host species
Rabbit -
Specificity
This reagent is not recommended for mouse or rat IHC-P and human ICC/IF.
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Tested applications
Suitable for: WB, IHC-Pmore details
Unsuitable for: ICC/IF or IP -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa whole cell lysate untreated or treated with human IFN gamma; A549, HEK-293T and MOLT-4 whole cell lysates; human fetal spleen, fetal kidney and thymus lysates; rat spleen and thymus lysates; mouse thymus lysate. IHC-P: human tonsil tissue and wild-type A549 cell pellet.
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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.
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 long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol (glycerin, glycerine), PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR20290 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab207728 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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WB |
1/1000. Detects a band of approximately 50 kDa (predicted molecular weight: 54 kDa).
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IHC-P |
1/100 - 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Notes |
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WB
1/1000. Detects a band of approximately 50 kDa (predicted molecular weight: 54 kDa). |
IHC-P
1/100 - 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Target
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Function
E3 ubiquitin-protein ligase whose activity is dependent on E2 enzymes, UBE2D1, UBE2D2, UBE2E1 and UBE2E2. Forms a ubiquitin ligase complex in cooperation with the E2 UBE2D2 that is used not only for the ubiquitination of USP4 and IKBKB but also for its self-ubiquitination. Component of cullin-RING-based SCF (SKP1-CUL1-F-box protein) E3 ubiquitin-protein ligase complexes such as SCF(SKP2)-like complexes. A TRIM21-containing SCF(SKP2)-like complex is shown to mediate ubiquitination of CDKN1B ('Thr-187' phosphorylated-form), thereby promoting its degradation by the proteasome. Monoubiquitinates IKBKB that will negatively regulates Tax-induced NF-kappa-B signaling. Negatively regulates IFN-beta production post-pathogen recognition by polyubiquitin-mediated degradation of IRF3. Mediates the ubiquitin-mediated proteasomal degradation of IgG1 heavy chain, which is linked to the VCP-mediated ER-associated degradation (ERAD) pathway. Promotes IRF8 ubiquitination, which enhanced the ability of IRF8 to stimulate cytokine genes transcription in macrophages. Plays a role in the regulation of the cell cycle progression. Enhances the decapping activity of DCP2. Exists as a ribonucleoprotein particle present in all mammalian cells studied and composed of a single polypeptide and one of four small RNA molecules. At least two isoforms are present in nucleated and red blood cells, and tissue specific differences in RO/SSA proteins have been identified. The common feature of these proteins is their ability to bind HY RNAs.2. -
Tissue specificity
Isoforms 1 and 2 are expressed in fetal and adult heart and fetal lung. -
Pathway
Protein modification; protein ubiquitination. -
Sequence similarities
Belongs to the TRIM/RBCC family.
Contains 1 B box-type zinc finger.
Contains 1 B30.2/SPRY domain.
Contains 1 RING-type zinc finger. -
Domain
The coiled-coil is necessary for the cytoplasmic localization. The B30.2/SPRY domain is necessary for the cytoplasmic localization, the interaction with IRF3 and for the IRF3-driven interferon beta promoter activity. The RING-type zinc finger is necessary for ubiquitination and for the IRF3-driven interferon beta promoter activity. Interacts with SKP2 and CUL1 in a RING finger-independent manner. -
Post-translational
modificationsAutoubiquitinated; does not lead to its proteasomal degradation. Deubiquitinated by USP4; leading to its stabilization. -
Cellular localization
Cytoplasm. Nucleus. Cytoplasm > P-body. Enters the nucleus upon exposure to nitric oxide. Localizes to small dot- or rod-like structures in the cytoplasm, called cytoplasmic bodies (P-body) that are located underneath the plasma membrane and also diffusely in the cytoplasm and are highly motil in cells. Cytoplasmic bodies are located along the microtubules and do not share the same cytoplasmic bodies with TRIM5. Colocalizes with DCP2 in P-body. - Information by UniProt
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Database links
- Entrez Gene: 6737 Human
- Entrez Gene: 20821 Mouse
- Entrez Gene: 308901 Rat
- Omim: 109092 Human
- SwissProt: P19474 Human
- SwissProt: Q62191 Mouse
- Unigene: 532357 Human
- Unigene: 321227 Mouse
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Alternative names
- 52 kDa ribonucleoprotein autoantigen Ro/SS-A antibody
- 52 kDa Ro protein antibody
- 52kD Ro/SSA autoantigen antibody
see all
Images
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Immunohistochemical analysis of paraffin-embedded A: Wild-type A549 (Human lung carcinoma epithelial cell) cell pellet, B: TRIM21 knockout A549 (ab267080) cell pellet labelling TRIM21/SS-A with ab207728 at 1/500 dilution (1.212 μg/ml) followed by LeicaDS9800 (Bond™ Polymer Refine Detection) secondary antibody at a ready to use concentration. Positive staining on (A) wild-type A549 cell pellet, no staining on (B) TRIM21 knockout A549 (ab267080) cell pellet. The section was incubated with ab207728 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
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Immunohistochemistry analysis of paraffin-embedded human tonsil tissue sections labelling TRIM21/SS-A with ab207728 at 1/100 dilution. The section was incubated with ab207728 for 10 mins at room temperature. Ready to use Leica DS9800 (Bond™ Polymer Refine Detection) was used as the secondary antibody. Sections were counterstained with Hematoxylin. Antigen retrieval was heat mediated with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 minutes.
Positive staining on human tonsil. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. -
All lanes : Anti-TRIM21/SS-A antibody [EPR20290] (ab207728) at 1/1000 dilution
Lane 1 : Wild-type A549 (Human lung carcinoma cell line) whole cell lysate
Lane 2 : TRIM21 knockout A549 (Human lung carcinoma cell line) whole cell lysate
Lane 3 : HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
Lane 4 : MOLT-4 (Human lymphoblastic leukemia 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: 54 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?Lanes 1-4: Merged signal (red and green). Green - ab207728 observed at 50 kDa. Red - loading control ab8245 observed at 36 kDa.
ab207728 Anti-TRIM21/SS-A antibody [EPR20290] was shown to specifically react with TRIM21/SS-A in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267024 (knockout cell lysate ab257766) was used. Wild-type and TRIM21/SS-A knockout samples were subjected to SDS-PAGE. ab207728 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-TRIM21/SS-A antibody [EPR20290] (ab207728) at 1/500 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : TRIM21 knockout A549 cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : MOLT-4 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: 54 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?Lanes 1-4: Merged signal (red and green). Green - ab207728 observed at 50 kDa. Red - loading control ab8245 observed at 36 kDa.
ab207728 Anti-TRIM21/SS-A antibody [EPR20290] was shown to specifically react with TRIM21/SS-A in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267025 (knockout cell lysate ab257767) was used. Wild-type and TRIM21/SS-A knockout samples were subjected to SDS-PAGE. ab207728 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 500 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-TRIM21/SS-A antibody [EPR20290] (ab207728) at 1/1000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : TRIM21 knockout HAP1 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lane 4 : MOLT-4 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 54 kDaLanes 1 - 4: Merged signal (red and green). Green - ab207728 observed at 50 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab207728 was shown to specifically react with in wild-type HAP1 cells as signal was lost in TRIM21 knockout cells. Wild-type and TRIM21 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% NF Milk. Ab207728 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 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.
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All lanes : Anti-TRIM21/SS-A antibody [EPR20290] (ab207728) at 1/1000 dilution
Lane 1 : Untreated HeLa (human epithelial cell line from cervix adenocarcinoma), whole cell lysate
Lane 2 : HeLa (human epithelial cell line from cervix adenocarcinoma) treated with 10 ng/ml human interferon-a (ab48750) for 16 hours
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 54 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
The level of TRIM21 expression can be elevated by IFN alpha treatment (PMID: 18071879).
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All lanes : Anti-TRIM21/SS-A antibody [EPR20290] (ab207728) at 1/1000 dilution
Lane 1 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen), whole cell lysate
Lane 2 : MOLT-4 (human lymphoblastic leukemia cell line), whole cell lysate
Lane 3 : Human fetal spleen lysate
Lane 4 : Human fetal kidney lysate
Lane 5 : Human thymus lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 54 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-TRIM21/SS-A antibody [EPR20290] (ab207728) at 1/1000 dilution
Lane 1 : Rat spleen lysate
Lane 2 : Rat thymus lysate
Lane 3 : Mouse thymus lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 54 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?Exposure times: Lane 1-2: 30 seconds; Lane 3: 3 minutes.
Blocking/Dilution buffer: 5% NFDM/TBST.
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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SDS download
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Datasheet download
Certificate of Compliance
References (5)
ab207728 has been referenced in 5 publications.
- Wang F et al. The Ubiquitin E3 Ligase TRIM21 Promotes Hepatocarcinogenesis by Suppressing the p62-Keap1-Nrf2 Antioxidant Pathway. Cell Mol Gastroenterol Hepatol 11:1369-1385 (2021). PubMed: 33482392
- Albecka A et al. A functional assay for serum detection of antibodies against SARS-CoV-2 nucleoprotein. EMBO J 40:e108588 (2021). PubMed: 34323299
- Zhou J et al. Identification of chemoresistance-related mRNAs based on gemcitabine-resistant pancreatic cancer cell lines. Cancer Med 9:1115-1130 (2020). PubMed: 31823522
- Chen J et al. Single-cell transcriptome and antigen-immunoglobin analysis reveals the diversity of B cells in non-small cell lung cancer. Genome Biol 21:152 (2020). PubMed: 32580738
- Hos NJ et al. TRIM21 Is Targeted for Chaperone-Mediated Autophagy during Salmonella Typhimurium Infection. J Immunol 205:2456-2467 (2020). PubMed: 32948684