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Applications: Useful for screening inhibitors of HSP90β and for HSP90β binding assays
Contraindications: Avoid DMSO concentrations above 0.5%. Avoid green and blue dyes that absorb light in the Alphascreen® signal emission range (520-620 nm), such as Trypan Blue. Avoid the use of the potent singlet oxygen quenchers such as sodium azide (NaN3) or metal ions (Fe2+, Fe3+, Cu2+, Zn2+ and Ni2+). The presence of >1% RPMI 1640 culture medium leads to a signal reduction due to the presence of excess biotin and iron in this medium. MEM, which lacks these components, does not affect Alphascreen® assays.
Description: The HSP90β (C-terminal) Inhibitor Screening Assay Kit is designed to measure the inhibition of HSP90β binding to its protein target PPID (also known as Cyclophilin D). With this kit, only three simple steps on a microtiter plate are required. First, a sample containing HSP90β, PPID, and an inhibitor of choice is incubated for thirty minutes. Next, acceptor beads are added, then donor beads, followed by reading the Alpha-counts.
Storage Stability: Stable at least 6 months from date of receipt, when stored as directed. Kit components require different storage conditions. Be sure to store each component at the proper temperature upon arrival.
Supplied As: Kit comes in a convenient AlphaLISA® format, with enough HSP90β (527-724), assay buffer, detection buffer, and purified GST-tagged PPID to perform a total of 384 enzyme reactions.
Uniprot: P08238
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: Allan, R.K. et al. J. Biol.Chem 2006 281(11): 7161-71.
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