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XIAP (X-linked inhibitor of apoptosis) belongs to the IAP (inhibitors of apoptosis) family, which is characterized by one or more BIR (baculovirus IAP repeat) domains. These proteins have multiple biological functions that include binding and inhibiting caspases, regulating cell cycle progression, and modulating receptor-mediated signal transduction. XIAP functions by direct binding and inhibition of several caspases, including caspase-3, caspase-7 and caspase-9.
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Western blot analysis of Jurkat (1), Raji (2), HeLa (3), WR19L (4) and PC12 (5) cells, probed with XIAP mAb (2F1).
XIAP ubiquitinates HIF1α. (A) U2OS cells were transfected with siRNA oligonucleotides and His-tagged ubiquitin as indicated. Lysates were prepared and precipitated using nickel-conjugated beads followed by western blotting using the indicated antibodies. (B) HA-tagged plasmid encoding HIF1α was coexpressed in HEK293 cells with His-tagged ubiquitin (Ub), FLAG-XIAP (X) or FLAG-VHL (V) as indicated. Lysates were prepared 48 h post-transfection and ubiquitinated material was recovered by incubation with nickel-conjugated beads and analysed by western blotting. (C) Biotinylated-HIF1α was coexpressed in HEK293 cells with FLAG-tagged XIAP. HIF1α was recovered from lysates by incubation with streptavidin-coated beads and analysed using antibodies directed against specific ubiquitin chains. (D) U2OS cells were pre-treated with 2μM NSC697923 for 30 min before being exposed to 1% O2 for 3 h. Cells were lysed in the presence of UBQ-TUBEs. Ubiquitin conjugates were purified using NiNTA agarose beads and analysed by immunoblotting using the indicated antibodies. (E) U2OS cells were pre-treated with 2 μM NSC697923 for 30 min before being exposed to 1% O2 for 3 h. Cells were lysed in the presence of TAB2-TUBEs or TAB2-AA-TUBEs as indicated. Proteins conjugated to Lys63-linked ubiquitin chains were purified using Ni-NTA agarose beads and analysed by immunoblot.
Image collected and cropped by CiteAb under a CC-BY license from the following publication: XIAP upregulates expression of HIF target genes by targeting HIF1α for Lys63-linked polyubiquitination. Nucleic Acids Res (2017)
| Regulatory Status |
RUO – Research Use Only |
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Related Products
| Application | ELISA, IHC, WB |
|---|---|
| Host | Goat |
| Species Reactivity | Mouse |