Human X-linked inhibitor of apoptosis (XIAP), also known as IAP-like protein, API3, X-linked IAP, and BIRC4 (GenBank Accession No. NM_001167), a.a. 2-end, with N-terminal FLAG-tag, MW= 57 kDa, expressed in a Baculovirus infected Sf9 cell expression system.

P98170

X-linked inhibitor of apoptosis, XIAP, IAP-like protein, API3, X-linked IAP, BIRC4

Ligase Assay Conditions:
50 µl of reaction mix (4 mM ATP, 5 mM MgCl₂, 1 mM DTT, E1 (0.5 µg), E2 (2.8 µg), Biotin-Ubiquitin (0.5 µg) and XIAP (0-1.25 µg) in 50 mM Tris pH 7.4) is incubated for 30 min at 37°C. Transfer the reaction mixture to a streptavidin-coated plate and incubate for 30 min at 37°C. Add antibody against XIAP, incubate 1 hr. Then, add secondary HRP-labeled antibody and incubate 30 min. Finally, add HRP chemiluminescent substrates and read luminescence.

Bir Domain Assay Conditions:
50 mM Tris-HCl, pH 7.4, 25 mM NaCl, 0.05% Tween-20, 0.01% BSA, 0.5 nM FAM-SMAC peptide. Incubate 30 minutes at room temperature. Fluorescence polarization is measured at 475 nm/528 nm of exc/em wavelength.

40 mM Tris-HCl, pH 8.0, 110 mM NaCl, 2.2 mM KCl, 80 μg/ml FLAG peptide, 20% glycerol, and 3 mM DTT.

Aqueous buffer solution

>6 months at -80°C.

Useful as an Ubiquitin ligase in conjunction with E1 and E2 for the study of enzyme kinetics. Ideal for Bir Domain binding and inhibition studies of Caspases 3 7, and 9. Also useful for screening inhibitors, and selectivity profiling.

1. Gu L. et al., Cancer Cell 15 (5), 363-375 (2009).
2. Deveraux Q.L. et al., Nature 388 (6639), 300-304 (1997).

Avoid freeze/thaw cycles

Ubiquitin E3 ligase

Recombinant Protein