HSP90α (C-Terminal Domain) TR-FRET Kit

Catalog #
50261
$1,125 *
Size: 384 reactions
Qty
*US Pricing only. For international pricing, please contact your local distributor.
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Description

The HSP90α (C-Terminal Domain) TR-FRET Assay Kit is designed to measure binding activity of HSP90α (heat shock protein 90 kDa alpha) to its target, PPID (peptidylprolyl isomerase D) for screening and profiling applications using TR-FRET (Time-Resolved Fluorescence Resonance Energy Transfer). It utilizes Terbium-labeled donor and a dye-labeled acceptor to complete the TR-FRET pairing. The HSP90α (C-Terminal Domain) TR-FRET Assay Kit comes in a convenient 384-well format, with enough purified HSP90α (C-Terminal) amino acids 535-732), PPID, Tb-Labeled Donor and Dye-Labeled Acceptor and assay buffer for 384 reactions.

Figure 1: Illustration of the assay principle.
A sample containing terbium-labeled donor, dye-labeled acceptor, HSP90α (C-Terminal), PPID, and an inhibitor is incubated. The fluorescence intensity is then measured using a fluorescence reader. In the presence of binding of HSP90α (C-Terminal) to PPID, energy transfer occurs due to the proximity of the donor and acceptor. Disruption of the binding results in decrease of energy transfer. Fluorescence intensity at λ=665 nm corresponds directly to the binding of HSP90α (C-Terminal) to PPID.

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Synonyms
HSP90a, Heat Shock Protein 90a, HSP90alpha, HSP90α, HTRF
Product Info
Storage and Usage
Citations
Assay Kit Format
TR-FRET
Materials Required But Not Supplied
  • Adjustable micropipettor and sterile tips
  • Fluorescence plate reader capable of measurement TR-FRET
Format
Catalog # Name Amount Storage
50316 HSP90α (C-terminal), Biotin-Labeled, His-Tag, Avi-Tag* 10 µg -80°C
71095 PPID (CYP-40), GST-Tag 10 µg -80°C
  Tb-Labeled Donor 2 x 10 µl -20°C
  Dye-Labeled Acceptor 2 x 10 µl -20°C
50324 3x HSP90 Assay Buffer 2 4 ml -20°C
79969 White 384-well microtiter plate 1 Room
Temp

* The concentration of protein is lot-specific and will be indicated on the tube containing the protein.

UniProt #
P07900
Background

HSP90 (heat shock protein 90 kDa) is a member of the HSP family of proteins. HSP proteins are chaperone proteins, involved in aiding proteins to fold correctly and resist heat related stress, cell signaling by acting on hormone receptors and kinases, cytoskeleton organization, cell cycle and differentiation by acting on CDK4 (cyclin dependent kinase 4), Wee1 and CDK11p110, among others. HSP90 has 3 family members, which differ in their cellular localization. HSP90A is a cytosolic protein and has two inducible isoforms (HSP90α1 and α2) and one that is constitutively expressed (HSP90β). HSP90B (endoplasmin) is found in the endoplasmic reticulum, and TRAP (TNF receptor-associated protein 1) in the mitochondria. They are composed of four domains: N-terminal domain (NTD), linker, middle domain (MD) and a C-terminal domain (CTD). The NTD has a high affinity ATP binding site, while the CTD is mainly involved in target binding, dimerization and localization. HSP90α is regulated at the transcription level, being induced at high temperatures, by post-translational modifications such as phosphorylation, and by co-chaperones. HSP90 can lead to cancer progression, by participating in the stabilization of several oncogenes and proteins involved in angiogenesis, inflammation, tumor suppression and metastasis. HSP90 has thus become an attractive therapeutic target for cancer therapy. In addition to cancer, HSP90 can also play a role in Alzheimer’s disease, Parkinson disease and viral infection. It is clear from its multiple and crucial functions that the development of inhibitors targeting HSP90 can prove beneficial for the treatment of several debilitating and fatal diseases.

References

Allan R.K., et al., 2006 J. Biol. Chem. 281(11): 7161-71.
Hoter A., et al., 2018 Int J Mol Sci. 19(9):2560