Overview of Enzyme - IMAP Assays
Based on the specific, high-affinity interaction of phospho groups with trivalent metal-containing nanoparticles (beads), IMAP is a generic, non-antibody-based platform to assess kinase, phosphatase, and phosphodiesterase activity. An enzyme reaction is performed using fluorescently labeled substrate. Addition of the IMAP Binding System stops the enzyme reaction and initiates binding of the beads to phosphorylated substrates. Binding of the substrate to the beads, which correlates to enzyme activity, can be detected using either FP or TR-FRET as a readout.
IMAP Progressive Binding System
IMAP Substrates
Latest Resources
Data of IMAP Assays
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Sample IMAP FP Data: ROCKII Dilution Curve
Enzyme dilution curves for ROCKII in BSA and in Tween IMAP Reaction Buffer using the Progressive Binding System. Reaction conditions: ROCKII kinase (Upstate: 14-451) as indicated, 100 nM FAM-S6 derived substrate (5FAM-AKRRRLSSLRA-COOH, R7184), 100 µM ATP. IMAP Binding Solution conditions: 100% Binding Buffer A, Progressive Binding Reagent 1:400.
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FP vs. TR-FRET Detection
Akt inhibition with staurosporine: Comparison of IMAP TR-FRET (top) and IMAP FP (bottom) detection. Reaction conditions: Akt kinase (0.1u/ml, Upstate: 14-276), 300 nM, 1µM, 3µM FAM-Crosstide (5FAM-GRPRTSSFAEG-COOH, R7110), 10 µM ATP, Staurosporine as indicated. IMAP Binding Solution: 40% Binding Buffer A, 60% Binding Buffer B, Progressive Binding Reagent 1:400.
Technology of Enzyme - IMAP Assays
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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 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.
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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 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.
Resources of Enzyme - IMAP Assays
eBook
Optimized Assays for Breakthrough Research
Optimized Assays for Breakthrough Research
Biology research has been greatly simplified by the availability of standardized assay kits, instruments, and assay protocols from commercial manufacturers.
Scientific Poster
Multiplexing and Cascade Assays with the IMAP Fluorescence Polarization Platform
Multiplexing and Cascade Assays with the IMAP Fluorescence Polarization Platform
This document highlights Multiplexing with IMAP Progressive Binding System, and cascade, or coupled, IMAP assays, enabling the HTS researcher to optimize their assay for each applica…
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Scientific Poster
Multiplex Assays in IMAP using the Progressive Binding System
Multiplex Assays in IMAP using the Progressive Binding System
The IMAP fluorescence polarization (FP) assay platform is a generic, homogeneous system applicable to a variety of enzymes, including protein kinases. IMAP is based on the high affin…
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Scientific Poster
Fluorescence Polarization Assays of Proteases with the IMAP Platform
Fluorescence Polarization Assays of Proteases with the IMAP Platform
Explore this document to know more about IMAP fluorescence polarization assay platform which is a non-antibody system applicable to wide variety of protein kinases.
Data Sheet
IMAP Technology for Kinases, Phosphatases and Phosphodiesterases
IMAP Technology for Kinases, Phosphatases and Phosphodiesterases
Get information about proprietary IMAP Technology which provides a non-radioactive, homogeneous assay for kinase activity.
Application Note
IMAP FP kinase assays on the SpectraMax M5 Multi-Mode Microplate Reader
IMAP FP kinase assays on the SpectraMax M5 Multi-Mode Microplate Reader
Protein kinases are central to the regulation of many cellular processes. In recent years they have emerged as one of the most important classes of drug targets for cancer and many…
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Application Note
IMAP phosphodiesterase assays on SpectraMax Multi-Mode Microplate Readers
IMAP phosphodiesterase assays on SpectraMax Multi-Mode Microplate Readers
Cyclic nucleotide phosphodiesterases (PDEs) are a group of enzymes that degrade the phosphodiester bond of cAMP and cGMP, second messengers that are involved in a variety of biological…
Compatible Products & Services of Enzyme - IMAP Assays
Featured Applications
IMAP FP generic kinase and phosphatase assays
IMAP principle using FP readout: Binding Solution is added after the kinase reaction using a…
IMAP FP generic phosphodiesterase assays
IMAP principle using FP readout: Binding Solution is added after the phosphodiesterase reaction…
IMAP TR-FRET generic kinase and phosphatase assays
IMAP principle using TR-FRET readout: Binding Solution is added after the kinase reaction using a…
IMAP TR-FRET generic phosphodiesterase assays
IMAP principle using TR-FRET readout: Binding Solution is added after the phosphodiesterase…
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