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  1. Link

    trim21ss-a-antibody-epr20290-ab207728.pdf

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Epigenetics and Nuclear Signaling DNA / RNA RNA Processing
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Validated using a knockout cell lineRecombinantRabMAb

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

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  • SDS
  • Certificate of Compliance
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRIM21/SS-A antibody [EPR20290] (ab207728)
  • Western blot - Anti-TRIM21/SS-A antibody [EPR20290] (ab207728)
  • Western blot - Anti-TRIM21/SS-A antibody [EPR20290] (ab207728)
  • Western blot - Anti-TRIM21/SS-A antibody [EPR20290] (ab207728)
  • Western blot - Anti-TRIM21/SS-A antibody [EPR20290] (ab207728)
  • Western blot - Anti-TRIM21/SS-A antibody [EPR20290] (ab207728)
  • Western blot - Anti-TRIM21/SS-A antibody [EPR20290] (ab207728)
  • Immunoprecipitation - Anti-TRIM21/SS-A antibody [EPR20290] (ab207728)
  • 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, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Recombinant Human TRIM21/SS-A protein (Tagged) (ab114611)
Primary
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Anti-Alpha-synuclein antibody [MJFR1] - BSA and Azide free (ab209420)
Knockout
Product image
Human TRIM21 (SS-A) knockout A549 cell lysate (ab257766)

View more associated products

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
  • Specificity

    This reagent is not recommended for mouse or rat IHC-P.

  • Tested applications

    Suitable for: WB, IHC-P, IPmore details
    Unsuitable for: ICC/IF
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • 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. IP: 293T (human embryonic kidney epithelial cell) whole cell lysate. IHC-P: human tonsil tissue.
  • 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.

Properties

  • 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...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR20290
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • RNA Processing
    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • RNA Processing
    • Splicing
    • Epigenetics and Nuclear Signaling
    • Chromatin Binding Proteins
    • DNA / RNA binding
    • Immunology
    • Immune System Diseases
    • Antiviral Signaling

Associated products

  • Alternative Versions

    • Anti-TRIM21/SS-A antibody [EPR20290] - BSA and Azide free (ab232549)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human TRIM21 (SS-A) knockout A549 cell line (ab267024)
    • Human TRIM21 (SS-A) knockout A549 cell line (ab267025)
  • KO cell lysates

    • Human TRIM21 (SS-A) knockout A549 cell lysate (ab257766)
    • Human TRIM21 (SS-A) knockout A549 cell lysate (ab257767)
  • Recombinant Protein

    • Recombinant Human TRIM21/SS-A protein (Tagged) (ab114611)
  • Related Products

    • Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
    • Recombinant human Interferon alpha 1 protein (Active) (ab48750)

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
WB
1/1000. Detects a band of approximately 50 kDa (predicted molecular weight: 54 kDa).
IHC-P
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP
1/30.
Notes
WB
1/1000. Detects a band of approximately 50 kDa (predicted molecular weight: 54 kDa).
IHC-P
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP
1/30.
Application notes
Is unsuitable for ICC/IF.

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.
  • Target information above from: UniProt accession P19474 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 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
    • 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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRIM21/SS-A antibody [EPR20290] (ab207728)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRIM21/SS-A antibody [EPR20290] (ab207728)

      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.

    • 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.

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

      TRIM21/SS-A was immunoprecipitated from 293T (human embryonic kidney epithelial cell) whole cell lysate 10 μg with ab207728 at 1/30 dilution.

      Western blot was performed on the immunoprecipitate using ab207728 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

      Lane 1: 293T (human embryonic kidney epithelial cell) whole cell lysate 10 μg.

      Lane 2: 293T whole cell lysate.

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab207728 in 293T whole cell lysate.

      Blocking and dilution buffer and concentration : 5% NFDM/TBST.

      Observed MW : 54 kDA.

      Exposure time: 3 minutes.

    • 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

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (4)

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

    ab207728 has been referenced in 4 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
    • 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

    Customer reviews and Q&As

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