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    sting-antibody-epr13130-ab181125.pdf

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Immunology Innate Immunity Cytokines Interferons
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-STING antibody [EPR13130] (ab181125)

  • Datasheet
  • SDS
  • Certificate of Compliance
Reviews (1) Submit a question References (11)

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Western blot - Anti-STING antibody [EPR13130] (ab181125)
  • Western blot - Anti-STING antibody [EPR13130] (ab181125)
  • Immunocytochemistry/ Immunofluorescence - Anti-STING antibody [EPR13130] (ab181125)
  • Flow Cytometry (Intracellular) - Anti-STING antibody [EPR13130] (ab181125)
  • Western blot - Anti-STING antibody [EPR13130] (ab181125)
  • Western blot - Anti-STING antibody [EPR13130] (ab181125)
  • Flow Cytometry (Intracellular) - Anti-STING antibody [EPR13130] (ab181125)
  • Anti-STING antibody [EPR13130] (ab181125)

Key features and details

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

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 594 Alexa Fluor® 647 Carrier Free HRP PE

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Anti-TMS1/ASC antibody [EPR10402(B)] (ab151700)
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Alexa Fluor® 488 Anti-STING antibody [EPR13130] (ab198950)
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HRP Anti-STING antibody [EPR13130] (ab198951)

View more associated products

Overview

  • Product name

    Anti-STING antibody [EPR13130]
    See all STING primary antibodies
  • Description

    Rabbit monoclonal [EPR13130] to STING
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, ICC/IFmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

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

  • Positive control

    • THP1 or HACAT lysate, THP1 or HACAT cells.
  • 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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

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

    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, 0.05% BSA, 59% PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR13130
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Innate Immunity
    • Cytokines
    • Interferons
    • Signal Transduction
    • Adapters
    • Transmembrane
    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • NFkB pathway
    • Immunology
    • Immune System Diseases
    • Antiviral Signaling
    • Cancer
    • Signal transduction
    • Nuclear signaling
    • NFkB pathway

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-STING antibody [EPR13130] (ab198950)
    • HRP Anti-STING antibody [EPR13130] (ab198951)
    • Alexa Fluor® 647 Anti-STING antibody [EPR13130] (ab198952)
    • Alexa Fluor® 594 Anti-STING antibody [EPR13130] (ab207288)
    • PE Anti-STING antibody [EPR13130] (ab208874)
    • Anti-STING antibody [EPR13130] - BSA and Azide free (ab227128)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • THP-1 whole cell lysate (ab7913)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab181125 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)
1/10 - 1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

WB (1)
1/1000 - 1/2000. Detects a band of approximately 37 kDa.
ICC/IF
1/2000.

For unpurified, use 1/50 - 1/100.

Notes
Flow Cyt (Intra)
1/10 - 1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

WB
1/1000 - 1/2000. Detects a band of approximately 37 kDa.
ICC/IF
1/2000.

For unpurified, use 1/50 - 1/100.

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.
  • Target information above from: UniProt accession Q86WV6 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 340061 Human
    • Omim: 612374 Human
    • SwissProt: Q86WV6 Human
    • Unigene: 379754 Human
    • 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] (ab181125)
      Western blot - Anti-STING antibody [EPR13130] (ab181125)
      All lanes : Anti-STING antibody [EPR13130] (ab181125) at 1/1000 dilution

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

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

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



      False colour image of Western blot: Anti-STING antibody [EPR13130] staining at 1/1000 dilution, shown in green; loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) staining at 1/20000 dilution, shown in red. In Western blot, ab181125 was shown to bind specifically to STING. A band was observed at 37 kDa in wild-type THP-1 cell lysates with no signal observed at this size in TMEM173 knockout cell line ab270493 (knockout cell lysate ab270516). To generate this image, wild-type and TMEM173 knockout THP-1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

    • Western blot - Anti-STING antibody [EPR13130] (ab181125)
      Western blot - Anti-STING antibody [EPR13130] (ab181125)
      All lanes : Anti-STING antibody [EPR13130] (ab181125) at 1/10000 dilution (purified)

      Lane 1 : THP-1 cell lysate
      Lane 2 : HACAT cell lysate
      Lane 3 : human spleen lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

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



      Blocking buffer: 5% NFDM/TBST

      Dilution buffer: 5% NFDM/TBST

    • Immunocytochemistry/ Immunofluorescence - Anti-STING antibody [EPR13130] (ab181125)
      Immunocytochemistry/ Immunofluorescence - Anti-STING antibody [EPR13130] (ab181125)

      Immunocytochemistry/immunofluorescence analysis of THP-1 (human monocytic leukemia monocyte) cells labelling TMEM173 with ab181125 at 10 µg/mL. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 was used as the secondary antibody (green). Cells were counterstained with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red). Nuclear DNA was labelled with DAPI (blue).
      Confocal image showing cytoplasmic staining in THP-1 cells.

    • Flow Cytometry (Intracellular) - Anti-STING antibody [EPR13130] (ab181125)
      Flow Cytometry (Intracellular) - Anti-STING antibody [EPR13130] (ab181125)

      Overlay histogram showing THP-1 cells fixed in 4% PFA and stained with purified ab181125 at a dilution of 1 in 20 (red line). The secondary antibody used was FITC goat anti-rabbit at a dilution of 1 in 500. Rabbit monoclonal IgG was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line).

      Alexa Fluor® 488 (ab198950) and R-PE (ab208874) conjugated versions are available for this clone.

    • Western blot - Anti-STING antibody [EPR13130] (ab181125)
      Western blot - Anti-STING antibody [EPR13130] (ab181125)
      Anti-STING antibody [EPR13130] (ab181125) at 1/2000 dilution (purified) + HEK293 cell lysate at 20 µg

      Secondary
      HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

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



      Blocking buffer: 5% NFDM/TBST

      Dilution buffer: 5% NFDM/TBST

    • Western blot - Anti-STING antibody [EPR13130] (ab181125)
      Western blot - Anti-STING antibody [EPR13130] (ab181125)
      All lanes : Anti-STING antibody [EPR13130] (ab181125) at 1/2000 dilution (unpurified)

      Lane 1 : THP1 lysate
      Lane 2 : HACAT lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
    • Flow Cytometry (Intracellular) - Anti-STING antibody [EPR13130] (ab181125)
      Flow Cytometry (Intracellular) - Anti-STING antibody [EPR13130] (ab181125)

      Intracellular Flow Cytometry analysis of THP1 cells using unpurified ab181125at a 1/10 dilution (red) or a Rabbit monoclonal IgG (negative) (green). Goat anti rabbit IgG (FITC) secondary used at a 1/150 dilution. 

       

       

    • Anti-STING antibody [EPR13130] (ab181125)
      Anti-STING antibody [EPR13130] (ab181125)

    Protocols

    • Flow cytometry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    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 (11)

    Publishing research using ab181125? Please let us know so that we can cite the reference in this datasheet.

    ab181125 has been referenced in 11 publications.

    • Lubbers JM  et al. Association of homozygous variants of STING1 with outcome in human cervical cancer. Cancer Sci 112:61-71 (2021). PubMed: 33040406
    • Wu X  et al. Porcine circovirus type 2 infection attenuates the K63-linked ubiquitination of STING to inhibit IFN-ß induction via p38-MAPK pathway. Vet Microbiol 258:109098 (2021). PubMed: 33984793
    • Miglietta G  et al. G-quadruplex binders as cytostatic modulators of innate immune genes in cancer cells. Nucleic Acids Res 49:6673-6686 (2021). PubMed: 34139015
    • Li R  et al. Brucella spp. Omp25 Promotes Proteasome-Mediated cGAS Degradation to Attenuate IFN-ß Production. Front Microbiol 12:702881 (2021). PubMed: 34394047
    • Vogt D  et al. STING is dispensable during KSHV infection of primary endothelial cells. Virology 540:150-159 (2020). PubMed: 31928996
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    Western blot abreview for Anti-TMEM173 antibody [EPR13130]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HEK293)
    Gel Running Conditions
    Reduced Denaturing (4-12%)
    Loading amount
    50 µg
    Specification
    HEK293
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 10°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    DR. Robert Kupp

    Verified customer

    Submitted Oct 19 2020

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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