Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-STING antibody [EPR13130-55] (ab239074)

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Western blot - Anti-STING antibody [EPR13130-55] (ab239074)
  • Western blot - Anti-STING antibody [EPR13130-55] (ab239074)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STING antibody [EPR13130-55] (ab239074)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STING antibody [EPR13130-55] (ab239074)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STING antibody [EPR13130-55] (ab239074)
  • Immunocytochemistry/ Immunofluorescence - Anti-STING antibody [EPR13130-55] (ab239074)
  • Immunoprecipitation - Anti-STING antibody [EPR13130-55] (ab239074)
  • Flow Cytometry (Intracellular) - Anti-STING antibody [EPR13130-55] (ab239074)
  • Anti-STING antibody [EPR13130-55] (ab239074)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR13130-55] to STING
  • Suitable for: Flow Cyt (Intra), WB, ICC/IF, IP, IHC-P
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-STING antibody [EPR13130-55]
    See all STING primary antibodies

  • Description

    Rabbit monoclonal [EPR13130-55] to STING

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, ICC/IF, IP, IHC-Pmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

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

  • Positive control

    • WB: Wild-type THP-1, THP-1 and HT1080 whole cell lysates; Human tonsil lysate. IHC-P: Human tonsil, colon cancer and prostate hyperplasia tissues. ICC/IF: THP-1 cells. Flow Cyt (intra): THP-1 cells. IP: THP-1 whole cell lysate.

  • 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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab239074 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
Flow Cyt (Intra)
1/500.
WB
1/1000. Detects a band of approximately 36 kDa (predicted molecular weight: 42 kDa).
ICC/IF
1/100.
IP
1/500.
IHC-P
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Notes
Flow Cyt (Intra)
1/500.
WB
1/1000. Detects a band of approximately 36 kDa (predicted molecular weight: 42 kDa).
ICC/IF
1/100.
IP
1/500.
IHC-P
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target

  • Function

    Facilitator of innate immune signaling that promotes the production of type I interferon (IFN-alpha and IFN-beta). Innate immune response is triggered in response to non-CpG double-stranded DNA from viruses and bacteria delivered to the cytoplasm. Able to activate both NF-kappa-B and IRF3 transcription pathways to induce expression of type I interferon and exert a potent anti-viral state following expression. May be involved in translocon function, the translocon possibly being able to influence the induction of type I interferons. May be involved in transduction of apoptotic signals via its association with the major histocompatibility complex class II (MHC-II). Mediates death signaling via activation of the extracellular signal-regulated kinase (ERK) pathway.

  • Tissue specificity

    Ubiquitously expressed.

  • Sequence similarities

    Belongs to the TMEM173 family.

  • Post-translational
    modifications

    Phosphorylated on tyrosine residues upon MHC-II aggregation (By similarity). Phosphorylated on Ser-358 by TBK1, leading to activation and production of IFN-beta.
    Ubiquitinated. 'Lys-63'-linked ubiquitination mediated by TRIM56 at Lys-150 promotes homodimerization and recruitment of the antiviral kinase TBK1 and subsequent production of IFN-beta. 'Lys-48'-linked polyubiquitination at Lys-150 occurring after viral infection is mediated by RNF5 and leads to proteasomal degradation.

  • Cellular localization

    Endoplasmic reticulum membrane. Mitochondrion outer membrane. Cell membrane. Cytoplasm > perinuclear region. In response to double-stranded DNA stimulation, relocalizes to perinuclear region, where the kinase TBK1 is recruited.
  • Information by UniProt

  • Database links

    • Alternative names

      • endoplasmic reticulum IFN stimulator antibody
      • Endoplasmic reticulum interferon stimulator antibody
      • ERIS antibody
      • FLJ38577 antibody
      • hMITA antibody
      • hSTING antibody
      • Mediator of IRF3 activation antibody
      • MITA antibody
      • Mitochondrial mediator of IRF3 activation antibody
      • MPYS antibody
      • N terminal methionine proline tyrosine serine plasma membrane tetraspanner antibody
      • NET23 antibody
      • Stimulator of interferon genes antibody
      • Stimulator of interferon genes protein antibody
      • STING antibody
      • TM173_HUMAN antibody
      • Tmem173 antibody
      • Transmembrane protein 173 antibody
      see all

    Images

    • Western blot - Anti-STING antibody [EPR13130-55] (ab239074)
      Western blot - Anti-STING antibody [EPR13130-55] (ab239074)
      All lanes : Anti-STING antibody [EPR13130-55] (ab239074) at 1/1000 dilution

      Lane 1 : Wild-type THP-1 cell lysate
      Lane 2 : TMEM173 knockout THP-1 cell lysate
      Lane 3 : Human Tonsil tissue lysate
      Lane 4 : Human Thymus tissue lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 42 kDa
      Observed band size: 37 kDa why is the actual band size different from the predicted?



      Lanes 1 - 4: Merged signal (red and green). Green - ab239074 observed at 37 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

      ab239074 was shown to react with TMEM173 in wild-type THP-1 cells in Western blot with loss of signal observed in TMEM173 knockout cell line ab270493 (TMEM173 knockout cell lysae ab270516). Wild-type THP-1 and TMEM173 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab239074 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 h at room temperature before imaging.

    • Western blot - Anti-STING antibody [EPR13130-55] (ab239074)
      Western blot - Anti-STING antibody [EPR13130-55] (ab239074)
      All lanes : Anti-STING antibody [EPR13130-55] (ab239074) at 1/1000 dilution

      Lane 1 : THP-1 (human monocytic leukemia cell line) whole cell lysate
      Lane 2 : HT1080 (human fibrosarcoma cell line) whole cell lysate
      Lane 3 : Human tonsil lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 42 kDa
      Observed band size: 36 kDa why is the actual band size different from the predicted?


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STING antibody [EPR13130-55] (ab239074)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STING antibody [EPR13130-55] (ab239074)

      Immunohistochemical analysis of paraffin-embedded human prostate hyperplasia tissue labeling TMEM173 with ab239074 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining in basal cells of human prostate hyperplasia is observed. Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STING antibody [EPR13130-55] (ab239074)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STING antibody [EPR13130-55] (ab239074)

      Immunohistochemical analysis of paraffin-embedded human colon cancer tissue labeling TMEM173 with ab239074 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining in human colon cancer (PMID: 26748708) is observed. Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STING antibody [EPR13130-55] (ab239074)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STING antibody [EPR13130-55] (ab239074)

      Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling TMEM173 with ab239074 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining in human tonsil (PMID:28874664) is observed. Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunocytochemistry/ Immunofluorescence - Anti-STING antibody [EPR13130-55] (ab239074)
      Immunocytochemistry/ Immunofluorescence - Anti-STING antibody [EPR13130-55] (ab239074)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized THP-1 (human monocytic leukemia cell line) cells labeling TMEM173 with ab239074 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and membranous staining in THP-1 cell line is observed.

      The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    • Immunoprecipitation - Anti-STING antibody [EPR13130-55] (ab239074)
      Immunoprecipitation - Anti-STING antibody [EPR13130-55] (ab239074)

      TMEM173 was immunoprecipitated from 0.35 mg of THP-1 (human monocytic leukemia cell line) whole cell lysate with ab239074 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab239074 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

      Lane 1: THP-1 whole cell lysate 10 µg (Input). 

      Lane 2: ab239074 IP in THP-1 whole cell lysate. 

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab239074 in THP-1 whole cell lysate.

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

      Exposure time: 30 seconds.

    • Flow Cytometry (Intracellular) - Anti-STING antibody [EPR13130-55] (ab239074)
      Flow Cytometry (Intracellular) - Anti-STING antibody [EPR13130-55] (ab239074)

      Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized THP-1 (human monocytic leukemia cell line) cell line labeling TMEM173 with ab239074 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

    • Anti-STING antibody [EPR13130-55] (ab239074)
      Anti-STING antibody [EPR13130-55] (ab239074)

    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 (0)

    Publishing research using ab239074? Please let us know so that we can cite the reference in this datasheet.

    ab239074 has not yet been referenced specifically in any publications.

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