Overview

  • Product name

    Human TRAF2 ELISA kit
    See all TRAF2 kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    PC3 ext 5 2.5%
    Inter-assay
    Sample n Mean SD CV%
    PC3 ext 3 1.1%
  • Sample type

    Cell culture extracts
  • Assay type

    Sandwich
  • Sensitivity

    1.5 ng/ml
  • Range

    1.6 ng/ml - 100 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture extracts 95 78% - 110%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    TRAF2 in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of TRAF2 protein in Human cell culture extract samples.


    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Wash Buffer PT (ab206977) 1 x 20ml
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent CPI 1 x 6ml
    10X Human TRAF2 Capture Antibody 1 x 600µl
    10X Human TRAF2 Detector Antibody 1 x 600µl
    Human TRAF2 Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS 1 x 12ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

  • Function

    Regulates activation of NF-kappa-B and JNK and plays a central role in the regulation of cell survival and apoptosis. Required for normal antibody isotype switching from IgM to IgG. Has E3 ubiquitin-protein ligase activity and promotes 'Lys-63'-linked ubiquitination of target proteins, such as BIRC3, RIPK1 and TICAM1. Is an essential constituent of several E3 ubiquitin-protein ligase complexes, where it promotes the ubiquitination of target proteins by bringing them into contact with other E3 ubiquitin ligases. Regulates BIRC2 and BIRC3 protein levels by inhibiting their autoubiquitination and subsequent degradation; this does not depend on the TRAF2 RING-type zinc finger domain.
  • Pathway

    Protein modification; protein ubiquitination.
  • Sequence similarities

    Belongs to the TNF receptor-associated factor family. A subfamily.
    Contains 1 MATH domain.
    Contains 1 RING-type zinc finger.
    Contains 2 TRAF-type zinc fingers.
  • Domain

    The coiled coil domain mediates homo- and hetero-oligomerization.
    The MATH/TRAF domain binds to receptor cytoplasmic domains.
    The RING-type zinc finger domain is essential for E3 ubiquitin-protein ligase activity. It is not essential for the stabilization of BIRC2, or for the ubiquitination of RIPK1 in response to TNFR1 signaling.
  • Post-translational
    modifications

    Phosphorylated at several serine residues within the first 128 amino acid residues. Phosphorylated at Thr-117 in response to signaling via TNF and TNFRSF1A. Phosphorylation at Thr-117 is required for 'Lys-63'-linked polyubiquitination, but not for 'Lys-48'-linked polyubiquitination. Phosphorylation at Thr-117 is important for interaction with IKKA and IKKB, activation of IKK and subsequent activation of NF-kappa-B.
    Undergoes both 'Lys-48'-linked and 'Lys-63'-linked polyubiquitination. Polyubiquitinated via 'Lys-63'-linked ubiquitin in response to TNF signaling; this requires prior phosphorylation at Thr-117. 'Lys-63'-linked polyubiquitination promotes TRAF2-mediated activation of NF-kappa-B. Can be polyubiquitinated at several Lys residues via 'Lys-48'-linked ubiquitin chains in response to TNF signaling, leading to proteasomal degradation. Autoubiquitinated, leading to its subsequent proteasomal degradation. Polyubiquitinated by BIRC2 and SIAH2, leading to its subsequent proteasomal degradation. Deubiquitinated by CYLD, a protease that specifically cleaves 'Lys-63'-linked polyubiquitin chains.
  • Cellular localization

    Cytoplasm.
  • Information by UniProt
  • Alternative names

    • E3 ubiquitin-protein ligase TRAF2
    • MGC:45012
    • OTTHUMP00000022625
    • OTTHUMP00000064745
    • TNF receptor associated factor 2
    • TNF receptor-associated factor 2
    • TNF receptor-associated protein
    • TRAF 2
    • TRAF2
    • TRAF2_HUMAN
    • TRAP
    • TRAP 3
    • TRAP3
    • Tumor necrosis factor type 2 receptor associated protein 3
    • Tumor necrosis factor type 2 receptor-associated protein 3
    see all
  • Database links

Associated products

Applications

Our Abpromise guarantee covers the use of ab226893 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
Sandwich ELISA Use at an assay dependent concentration.

Images

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of TRAF2 were measured in duplicate and interpolated from the TRAF2 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean TRAF2 concentration was determined to be 57 ng/mL in HeLa, 61 ng/mL in MOLT-4, and 34 ng/ml in PC-3.

  • Interpolated concentrations of native TRAF2 in human and mouse extracts based on a 1,000 μg/mL extract load. The concentrations of native TRAF2 were measured in duplicate, interpolated from the human TRAF2 standard curve, and corrected for sample dilution. The mean TRAF2 concentrations are plotted (n=5).

Protocols

References

ab226893 has not yet been referenced specifically in any publications.

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