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.
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.
*The concentration of HSP90 and PPID are lot-specific and will be indicated on the tubes containing the protein
UniProt #
P07900
References
Allan, R.K. et al. J. Biol.Chem 2006. 281(11): 7161-71.
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.
Applications
Useful for screening inhibitors of HSP90α and for HSP90α binding assays
Shipping Temperature
-80°C
Notes
AlphaScreen® and AlphaLISA® are registered trademarks of PerkinElmer, Inc.
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.
Warnings
Avoid freeze/thaw cycles.
Disclaimers and Limitations
BPS Bioscience assay kits are validated using the listed components according to our specific protocol. Any deviations to kit components or protocols, such as using different proteins, cell/tissue lysates, or buffers (including using the same component from other commercial sources) will void any warranties on the performance of the assay kit and is not recommended.
End-users should immediately report any issues to [email protected] upon receipt of the kit, such as missing components, damaged vials, no dry ice, etc.