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Applications: Useful for screening inhibitors of HSP90α and for HSP90α binding assays
Contraindications: Only limited amounts of DMSO (< 1%) can be included as it has been shown to disrupt HSP90α:PPID interaction. 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 HSP90α (C-Terminal) Inhibitor Screening Assay Kit is designed to measure the inhibition of HSP90α 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 HSP90α, 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: The assay kit comes in a convenient AlphaLISA® format, with enough HSP90α (535-732), assay buffer, detection buffer, and purified GST-tagged PPID to perform a total of 384 enzyme reactions.
Uniprot: P07900
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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