Overview

  • Product name

    Human XIAP ELISA Kit
    See all XIAP kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Cell extract 6 2%
  • Sample type

    Cell culture extracts
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    114.3 pg/ml
  • Range

    0.47 ng/ml - 30 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture extracts 98.5 94.3% - 102.9%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    XIAP in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of XIAP protein in human cell lysate.


    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 Development Solution 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.


    XIAP plays a key role in various cellular processes, such as the regulation of cell proliferation, apoptosis, inflammation, proteasomal regulation, and immunity. One of XIAP’s predominant roles is the blocking of apoptosis through binding and direct inhibition of caspases. Increased expression of XIAP is a hallmark of some cancers including those of the lung and prostate.

  • 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 Human XIAP Capture Antibody 1 x 600µl
    10X Human XIAP Detector Antibody 1 x 600µl
    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 4BI 1 x 6ml
    Human XIAP Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS (ab193972) 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

    Apoptotic suppressor. Has E3 ubiquitin-protein ligase activity. Mediates the proteasomal degradation of target proteins, such as caspase-3, SMAC or AIFM1. Inhibitor of caspase-3, -7 and -9. Mediates activation of MAP3K7/TAK1, leading to the activation of NF-kappa-B.
  • Tissue specificity

    Ubiquitous, except peripheral blood leukocytes.
  • Involvement in disease

    Defects in XIAP are the cause of lymphoproliferative syndrome X-linked type 2 (XLP2) [MIM:300635]. XLP is a rare immunodeficiency characterized by extreme susceptibility to infection with Epstein-Barr virus (EBV). Symptoms include severe or fatal mononucleosis, acquired hypogammaglobulinemia, pancytopenia and malignant lymphoma.
  • Sequence similarities

    Belongs to the IAP family.
    Contains 3 BIR repeats.
    Contains 1 RING-type zinc finger.
  • Domain

    The first BIR domain is involved in interaction with TAB1/MAP3K7IP1 and is important for dimerization. The second BIR domain is sufficient to inhibit caspase-3 and caspase-7, while the third BIR is involved in caspase-9 inhibition. The interactions with SMAC and PRSS25 are mediated by the second and third BIR domains.
  • Post-translational
    modifications

    Ubiquitinated and degraded by the proteasome in apoptotic cells.
    Phosphorylation by PKB/AKT protects XIAP against ubiquitination and protects the protein against proteasomal degradation.
  • Cellular localization

    Cytoplasm.
  • Information by UniProt
  • Alternative names

    • AP 13
    • API3
    • Apoptosis Inhibitor 3
    • Baculoviral IAP repeat containing 4
    • Baculoviral IAP Repeat Containing Protein 4
    • Baculoviral IAP repeat-containing protein 4
    • BIRC 4
    • BIRC4
    • E3 ubiquitin-protein ligase XIAP
    • hIAP-3
    • hIAP3
    • HILP
    • IAP 3
    • IAP like protein
    • IAP-3
    • IAP-like protein
    • IAP3
    • ILP
    • ILP 1
    • ILP1
    • Inhibitor of apoptosis protein 3
    • Inhibitor of Apoptosis X Linked
    • Mammalian IAP Homologue A
    • MIHA
    • X linked IAP
    • X linked inhibitor of apoptosis
    • X linked inhibitor of apoptosis E3 ubiquitin protein ligase
    • X linked inhibitor of apoptosis protein
    • X-linked IAP
    • X-linked inhibitor of apoptosis protein
    • Xiap
    • XIAP_HUMAN
    • XLP2
    see all
  • Database links

Associated products

Applications

Our Abpromise guarantee covers the use of ab234576 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 XIAP were measured in duplicate and interpolated from the XIAP standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean XIAP concentration was determined to be 8.2 ng/mL in sample and 18.2 ng/mL in sample.

Protocols

References

ab234576 has not yet been referenced specifically in any publications.

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