Recombinant
RabMAb

Recombinant Anti-TIFA (phospho T9) antibody [EPR19853] (ab214815)

Overview

  • Product name

    Anti-TIFA (phospho T9) antibody [EPR19853]
    See all TIFA primary antibodies
  • Description

    Rabbit monoclonal [EPR19853] to TIFA (phospho T9)
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, Dot blot, IPmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human TIFA aa 1-100. The exact sequence is proprietary.
    Database link: Q96CG3

  • Positive control

    • WB: HeLa transfected with 3×Flag-tagged TIFA expression vector for 24 hours, and then infected with S. flexneri 2457T for 4 hours, whole cell lysate. HEK-293T transfected with GFP-tagged TIFA expression vector for 24 hours, and then infected with B. cenocepacia J2315 for 4 hours, whole cell lysate. IP: HEK-293T transfected with GFP-tagged TIFA expression vector for 24 hours, and then infected with B. cenocepacia J2315 for 4 hours whole cell lysate.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab214815 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. Predicted molecular weight: 21 kDa.
Dot blot 1/1000.
IP 1/30.

Target

  • Function

    Adapter protein which mediates the IRAK1 and TRAF6 interaction following IL-1 stimulation, resulting in the downstream activation of NF-kappa-B and AP-1 pathways. Induces the oligomerization and polyubiquitination of TRAF6, which leads to the activation of TAK1 and IKK through a proteasome-independent mechanism.
  • Sequence similarities

    Contains 1 FHA domain.
  • Information by UniProt
  • Database links

  • Alternative names

    • Putative MAPK activating protein PM14 antibody
    • Putative MAPK-activating protein PM14 antibody
    • Putative NF kappa B activating protein 20 antibody
    • Putative NF-kappa-B-activating protein 20 antibody
    • T2BP antibody
    • TIFA antibody
    • TIFA_HUMAN antibody
    • TRAF interacting protein with FHA domain containing protein A antibody
    • TRAF-interacting protein with FHA domain-containing protein A antibody
    • TRAF2 binding protein antibody
    • TRAF2-binding protein antibody
    see all

Images

  • All lanes : Anti-TIFA (phospho T9) antibody [EPR19853] (ab214815) at 1/1000 dilution

    Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) transfected with 3×Flag-tagged TIFA expression vector for 28 hours, whole cell lysate
    Lane 2 : HeLa transfected with 3×Flag-tagged TIFA T9A mutant expression vector for 28 hours, whole cell lysate
    Lane 3 : HeLa transfected with 3×Flag-tagged TIFA expression vector for 24 hours, and then infected with S. flexneri 2457T for 4 hours, whole cell lysate
    Lane 4 : HeLa transfected with 3×Flag-tagged TIFAT9A mutant expression vector for 24 hours and then infected with S. flexneri 2457T for 4 hours, whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/5000 dilution

    Predicted band size: 21 kDa
    Observed band size: 26 kDa
    why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% milk/TBST.

    The image was kindly provided by our collaborator Dr. Feng Shao, NIBS.

     Sflexneri infection induces an innate immune response (PMID: 28222186).

  • TIFA (phospho T9) was immunoprecipitated from 0.35 mg HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate with ab214815 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab214815 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5,000 dilution

    Lane 1: HEK-293T transfected with GFP-tagged TIFA expression vector for 24 hours, and then infected with B. cenocepacia J2315 for 4 hours whole cell lysate 10 µg (Input).
    Lane 2: ab214815 IP in HEK-293T transfected with GFP-tagged TIFA expression vector for 24 hours, and then infected with B. cenocepacia J2315 for 4 hours whole cell lysate (+).
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab214815 in HEK-293T transfected with GFP-tagged TIFA expression vector for 24 hours, and then infected with B. cenocepacia J2315 for 4 hours whole cell lysate (-).

    Blocking/Dilution buffer: 5% NFDM/TBST.
    Exposure time: 3 minutes.

    The cell lysates were kindly provided by our collaborator Dr. Feng Shao, NIBS.

  • All lanes : Anti-TIFA (phospho T9) antibody [EPR19853] (ab214815) at 1/5000 dilution

    Lane 1 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with GFP-tagged TIFA expression vector, whole cell lysate
    Lane 2 : HEK-293T transfected with GFP-tagged TIFA expression vector for 24 hours, and then infected with B. cenocepacia J2315 for 4 hours, whole cell 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: 21 kDa
    Observed band size: 47 kDa why is the actual band size different from the predicted?


    Exposure time: 103 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The band observed represents GFP-tagged TIFA pT9.

    The lysates were kindly provided by our collaborator Dr. Feng Shao, NIBS.

    B. cenocepacia J2315 infection elicits an inflammatory response. (PMID: 27133449)

     

  • Dot blot analysis of TIFA (phospho T9) labeled with ab214815 at 1/1000 dilution.

    Lane 1: TIFA (phospho T9) peptide (aa3-12).

    Lane 2: TIFA (phospho T9) peptide (aa5-15).

    Lane 3: TIFA non-phospho peptide (aa3-15).

    Secondary used was Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: 10 seconds.

References

This product has been referenced in:

See 1 Publication for this product

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