IMAP Kinase, Phosphatase, and Phosphodiesterase Assays


Homogeneous assay for accurate determination of kinase, phosphatase, and phosphodiesterase activities

IMAP® technology provides a homogeneous assay applicable to a wide variety of kinases without regard for substrate peptide sequence. The assay is a simple “mix-and-read” procedure that allows accurate determination of kinase, phosphatase, and phosphodiesterase (PDE) activity.

 

IMAP FP assay principle

Figure 1. IMAP FP generic kinase and phosphatase assays
IMAP principle using FP readout: Binding Solution is added after the kinase reaction using a fluorescently labeled peptide. The small phosphorylated fluorescent substrate binds to the large M(III)-based nanoparticles which reduces the rotational speed of the substrate and thus increases its polarization.

IMAP TR-FRET

Figure 2. IMAP TR-FRET generic kinase and phosphatase assays
IMAP principle using TR-FRET readout: Binding Solution is added after the kinase reaction using a fluorescently labeled peptide or protein. In this system, the nanoparticle is spiked with a Terbium (Tb)-Donor molecule. By binding to the spiked M(III)-based nanoparticles, the phosphorylated fluorescent substrate comes into close proximity with the Tb-Donor, which allows measurement of the TR-FRET between the Tb-Donor and the phosphorylated, fluorescent substrate.

IMAP PDE FP assay principle

Figure 3. IMAP FP generic phosphodiesterase assays
IMAP principle using FP readout: Binding Solution is added after the phosphodiesterase reaction using a fluorescently labeled substrate. The small phosphorylated fluorescent substrate binds to the large M(III)-based nanoparticles which reduces the rotational speed of the substrate and thus increases its polarization.

IMAP PDE TR-FRET assay principle

Figure 4. IMAP TR-FRET generic phosphodiesterase assays
IMAP principle using TR-FRET readout: Binding Solution is added after the phosphodiesterase reaction using a fluorescently labeled substrate. In this system, the nanoparticle is spiked with a Terbium (Tb)-Donor molecule. By binding to the spiked M(III)-based nanoparticles, the phosphorylated fluorescent substrate comes into close proximity with the Tb-Donor, which allows measurement of the TR-FRET between the Tb-Donor and the phosphorylated, fluorescent substrate.

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