Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-TRIM21/SS-A antibody [EPR20290] (ab207728)

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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
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • 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

  • Tested applications

    Suitable for: WBmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment within Human TRIM21/SS-A aa 1-250. The exact sequence is proprietary.
    Database link: P19474

  • 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.

  • General notes

    Previously labelled as TRIM21

    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.

    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

Applications

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
WB 1/1000. Detects a band of approximately 50 kDa (predicted molecular weight: 54 kDa).

Target

  • 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
    modifications

    Autoubiquitinated; 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

  • Database links

    see all

  • Alternative names

    • 52 kDa ribonucleoprotein autoantigen Ro/SS-A antibody
    • 52 kDa Ro protein antibody
    • 52kD Ro/SSA autoantigen antibody
    • Autoantigen Ro/SSA, 52-KD antibody
    • E3 ubiquitin-protein ligase TRIM21 antibody
    • RING finger protein 81 antibody
    • RNF81 antibody
    • Ro 52 antibody
    • Ro(SS-A) antibody
    • Ro52 antibody
    • RO52_HUMAN antibody
    • Sicca syndrome antigen A antibody
    • Sjoegren syndrome type A antigen antibody
    • Sjogren syndrome antigen A1 antibody
    • Sjogren syndrome type A antigen antibody
    • SS-A antibody
    • SSA antibody
    • SSA1: Sjogren syndrome antigen A1 (52kDa ribonucleoprotein autoantigen SS-A/Ro) antibody
    • TRIM21 antibody
    • Tripartite motif protein TRIM21 antibody
    • Tripartite motif-containing 21 antibody
    • Tripartite motif-containing protein 21 antibody
    see all

Images

  • Western blot - Anti-TRIM21/SS-A antibody [EPR20290] (ab207728)
    Western blot - Anti-TRIM21/SS-A antibody [EPR20290] (ab207728)
    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.

  • Western blot - Anti-TRIM21/SS-A antibody [EPR20290] (ab207728)
    Western blot - Anti-TRIM21/SS-A antibody [EPR20290] (ab207728)
    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.

  • Western blot - Anti-TRIM21/SS-A antibody [EPR20290] (ab207728)
    Western blot - Anti-TRIM21/SS-A antibody [EPR20290] (ab207728)
    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 kDa



    Lanes 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.

  • Western blot - Anti-TRIM21/SS-A antibody [EPR20290] (ab207728)
    Western blot - Anti-TRIM21/SS-A antibody [EPR20290] (ab207728)
    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 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The level of TRIM21 expression can be elevated by IFN alpha treatment (PMID: 18071879).

  • Western blot - Anti-TRIM21/SS-A antibody [EPR20290] (ab207728)
    Western blot - Anti-TRIM21/SS-A antibody [EPR20290] (ab207728)
    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 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-TRIM21/SS-A antibody [EPR20290] (ab207728)
    Western blot - Anti-TRIM21/SS-A antibody [EPR20290] (ab207728)
    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.

  • Anti-TRIM21/SS-A antibody [EPR20290] (ab207728)
    Anti-TRIM21/SS-A antibody [EPR20290] (ab207728)

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.

Click here to view the general protocols

References (2)

Publishing research using ab207728? Please let us know so that we can cite the reference in this datasheet.

ab207728 has been referenced in 2 publications.

  • 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

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