Cellomics HCS Reagent Kit for detection of immunophilin FK506-binding protein 52 (FKBP52).

normal FKBP52 expression	treat cells	increased FKBP52 expression	analyze	conclude

Description

The Cellomics Immunophilin FKBP52 Detection Kit is for simultaneous quantification of nuclear DNA content and FK506-binding protein 52 (FKBP52) (Figure 1). This kit allows direct measurements using a fixed end-point assay based on immunofluorescence detection in cells grown on standard high-density microplates. The DNA binding dye, DAPI, enables nuclear size and morphology determination and cell cycle phase identification by DNA content. The anti-FKBP52 antibody is specific with minimal cross-reactivity.

This kit detects inducible FKBP52 (also known as FKBP4, FKBP59, Hsp56, Hsp59) protein in the cell. FKBP52 is a large immunophilin that binds to the immunosuppressive drug FK506 and has peptidyl-prolyl cis-trans isomerase (PPIase) activity, which is inhibited by the binding of FK506. Immunophilins are enriched in the central and peripheral neurons and reports indicate that FKBP52 has neurotrophic activity. FKBP52 contains three tetratricopeptide repeat (TPR) domains that mediate protein-protein interactions with Hsp90 and a calmodulin-binding site in the C-terminal region. In addition to neuroprotection, FKBP52 is involved in the maturation and translocation of steroid hormone receptors, transcription regulation, copper transport and cardiotrophic effect.

This kit was optimized with the Thermo Scientific ArrayScan HCS Reader using the Compartmental Analysis BioApplication Software Module. Thus, automated plate-handling, focusing, cell image acquisition and processing, and data analysis and management are combined in one high-content screening (HCS) system. In addition to HCS instruments, cells labeled by the kit reagents can be viewed and analyzed by fluorescence and confocal microscopes.

Figure 1. Staining of FKBP52 and nuclei in A549 cells (20X objective). The cells were treated media only (non-treated) or with CuSO4 (1 mM), stained according to the kit protocol and imaged using the ArrayScan VTI HCS Reader.

Figure 2. Chemicals induce FKBP52 in A549 cells. Different chemicals (Left panel: CuSO4, Right panel: menadione and ZnCl2) were added to the cells at the indicated concentrations. Each data point represents 16 wells from three separate plates. FKBP52 was measured using the output parameter of cytoplasmic or nuclear fluorescence or the difference between cytoplasm and nucleus average fluorescence intensity (cyt-nuc diff) with Compartmental Analysis Bioapplication. Values are normalized with the maximum control value.

Kit Contents

Ordering Information

Screening size kits and components (bulk and custom) available with special pricing. Please call.

To order Cellomics HCS Reagent Kits, please call 800-874-3723 in the U.S.
or contact your local distributor of Thermo Scientific Products.

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