Human XIAP ELISA Kit is a single-wash 90-min SimpleStep ELISA® for the quantitative measurement of Human XIAP in Cell culture extracts samples.
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Key facts
- Detection method
- Colorimetric
- Sample types
- Cell culture extracts
- Assay type
- Sandwich (quantitative)
- Reactive species
- Human
- Range
- 0.47 - 30 ng/mL
- Assay time
- 1h 30m
- Sensitivity
- = 114.3 pg/mL
Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application sELISA | Reactivity Reacts | Dilution info - | Notes - |
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Target data
Function
Multi-functional protein which regulates not only caspases and apoptosis, but also modulates inflammatory signaling and immunity, copper homeostasis, mitogenic kinase signaling, cell proliferation, as well as cell invasion and metastasis (PubMed:11257230, PubMed:11257231, PubMed:11447297, PubMed:12121969, PubMed:12620238, PubMed:17560374, PubMed:17967870, PubMed:19473982, PubMed:20154138, PubMed:22103349, PubMed:9230442). Acts as a direct caspase inhibitor (PubMed:11257230, PubMed:11257231, PubMed:12620238). Directly bind to the active site pocket of CASP3 and CASP7 and obstructs substrate entry (PubMed:11257230, PubMed:11257231, PubMed:16352606, PubMed:16916640). Inactivates CASP9 by keeping it in a monomeric, inactive state (PubMed:12620238). Acts as an E3 ubiquitin-protein ligase regulating NF-kappa-B signaling and the target proteins for its E3 ubiquitin-protein ligase activity include: RIPK1, RIPK2, MAP3K2/MEKK2, DIABLO/SMAC, AIFM1, CCS, PTEN and BIRC5/survivin (PubMed:17560374, PubMed:17967870, PubMed:19473982, PubMed:20154138, PubMed:22103349, PubMed:22607974, PubMed:29452636, PubMed:30026309). Acts as an important regulator of innate immunity by mediating 'Lys-63'-linked polyubiquitination of RIPK2 downstream of NOD1 and NOD2, thereby transforming RIPK2 into a scaffolding protein for downstream effectors, ultimately leading to activation of the NF-kappa-B and MAP kinases signaling (PubMed:19667203, PubMed:22607974, PubMed:29452636, PubMed:30026309). 'Lys-63'-linked polyubiquitination of RIPK2 also promotes recruitment of the LUBAC complex to RIPK2 (PubMed:22607974, PubMed:29452636). Regulates the BMP signaling pathway and the SMAD and MAP3K7/TAK1 dependent pathways leading to NF-kappa-B and JNK activation (PubMed:17560374). Ubiquitination of CCS leads to enhancement of its chaperone activity toward its physiologic target, SOD1, rather than proteasomal degradation (PubMed:20154138). Ubiquitination of MAP3K2/MEKK2 and AIFM1 does not lead to proteasomal degradation (PubMed:17967870, PubMed:22103349). Plays a role in copper homeostasis by ubiquitinating COMMD1 and promoting its proteasomal degradation (PubMed:14685266). Can also function as E3 ubiquitin-protein ligase of the NEDD8 conjugation pathway, targeting effector caspases for neddylation and inactivation (PubMed:21145488). Ubiquitinates and therefore mediates the proteasomal degradation of BCL2 in response to apoptosis (PubMed:29020630). Protects cells from spontaneous formation of the ripoptosome, a large multi-protein complex that has the capability to kill cancer cells in a caspase-dependent and caspase-independent manner (PubMed:22095281). Suppresses ripoptosome formation by ubiquitinating RIPK1 and CASP8 (PubMed:22095281). Acts as a positive regulator of Wnt signaling and ubiquitinates TLE1, TLE2, TLE3, TLE4 and AES (PubMed:22304967). Ubiquitination of TLE3 results in inhibition of its interaction with TCF7L2/TCF4 thereby allowing efficient recruitment and binding of the transcriptional coactivator beta-catenin to TCF7L2/TCF4 that is required to initiate a Wnt-specific transcriptional program (PubMed:22304967).
Alternative names
API3, BIRC4, IAP3, XIAP, E3 ubiquitin-protein ligase XIAP, Baculoviral IAP repeat-containing protein 4, IAP-like protein, Inhibitor of apoptosis protein 3, RING-type E3 ubiquitin transferase XIAP, X-linked inhibitor of apoptosis protein, ILP, hILP, IAP-3, hIAP-3, hIAP3, X-linked IAP
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Human XIAP ELISA Kit is a single-wash 90-min SimpleStep ELISA® for the quantitative measurement of Human XIAP in Cell culture extracts samples.
Key facts
- Detection method
- Colorimetric
- Sample types
- Cell culture extracts
- Assay type
- Sandwich (quantitative)
- Reactive species
- Human
- Range
- 0.47 - 30 ng/mL
- Assay time
- 1h 30m
- Assay Platform
- Pre-coated microplate (12 x 8 well strips)
- Sensitivity
- = 114.3 pg/mL
Precision
Intra assay
| Sample | n | mean | SD | C.V. |
|---|---|---|---|---|
Sample Cell Extract | n 6 | mean - | SD - | C.V. 2 |
Recovery
Sample specific recovery
| Sample type | Average % | Range |
|---|---|---|
Sample type Cell culture extracts | Average % = 98.5 | Range 94.3 - 102.9 % |
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- +4°C
- Storage information
- +4°C
Notes
Human XIAP ELISA Kit (ab234576) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of XIAP protein in cell culture extracts. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human XIAP with 114.3 pg/ml sensitivity.
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:
- Single-wash protocol reduces assay time to 90 minutes or less
- High sensitivity, specificity and reproducibility from superior antibodies
- Fully validated in biological samples
- 96-wells plate breakable into 12 x 8 wells strips
A 384-well SimpleStep ELISA® microplate (Pre-coated 384 well Microplate SimpleStep ELISA® ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
XIAP also known as X-linked inhibitor of apoptosis protein is a member of the inhibitor of apoptosis protein (IAP) family. XIAP has a molecular mass of approximately 57 kDa. This protein inhibits apoptosis by binding to and inhibiting caspases particularly caspase-3 caspase-7 and caspase-9. XIAP expresses in a variety of tissues including the heart kidney and liver but also shows expression in cancer cells. Its role in inhibiting cell death makes it a significant focus for cancer research.
- Biological function summary
The function of XIAP involves regulation of apoptosis an important process in maintaining cellular homeostasis. It forms part of the apoptosome complex by interacting with caspases to prevent premature cell death. XIAP serves to protect cells from stress-induced apoptotic signals helping in survival under adverse conditions. This ability makes it a subject of interest in studying cancer survival mechanisms since overactive XIAP can allow cancer cells to avoid programmed cell death.
- Pathways
Apoptosis regulation and cell survival pathways prominently feature XIAP. Particularly XIAP plays a role in the intrinsic apoptotic pathway which is activated in response to internal stress signals. XIAP regulates this pathway via its interaction with caspase-9 and interactions with other proteins such as Smac/DIABLO which can antagonize XIAP's function. This interaction is key in balancing cell death and survival therefore highlighting its role in apoptosis control.
- Associated diseases and disorders
XIAP's dysregulation links to cancer and neurodegenerative disorders. Overexpression of XIAP can contribute to cancer by enabling cells to evade apoptosis leading to uncontrolled cell proliferation. Moreover studies have suggested a role in neurodegenerative diseases where alterations in XIAP may affect neuronal cell survival. In these contexts XIAP's interactions with proteins such as caspases and Smac/DIABLO surface as important in understanding how apoptosis evasion or enhancement contributes to disease pathology.
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2 product images
Sandwich ELISA - Human XIAP ELISA Kit (ab234576)
Example of human XIAP standard curve in 1X Cell Extraction Buffer PTR.
Background-subtracted data values (mean +/- SD) are graphed.
Sandwich ELISA - Human XIAP ELISA Kit (ab234576)
Interpolated concentrations of native XIAP in human cell extracts based on a 1,000 μg/mL and 250 μg/mL extract load for MCF-7 and Raji cell, respectively.
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.
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