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

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Data of IMAP Assays

  • Sample IMAP data: ROCKII dilution curve

    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.

  • 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.

    FP vs. TR-FRET detection

Technology of Enzyme - IMAP Assays

  • IMAP FP generic kinase and phosphatase assays

    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 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 TR-FRET generic kinase and phosphatase assays

    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 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.

Ordering Options of Enzyme - IMAP Assays

IMAP Assay Kit

IMAP Evaluation Demo Kit - PN: R8166

IMAP Evaluation Demo Kit
Optimized 800 data point IMAP kit which includes calibrators (phosphorylated and un-phosphorylated peptides) to run a calibration curve for either FP or TR-FRET detection

#R8166

$822.00 USD

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IMAP TR-FRET Evaluation Kit (with Progressive Binding System) - PN: R8161

IMAP TR-FRET Evaluation Kit with Progressive Binding System
Optimized 800 data point protocol to run IMAP with the Progressive Binding System; Additional kinase and substrate are required

#R8161

$822.00 USD

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IMAP FP PDE Evaluation Kit - PN: R8175

IMAP FP PDE Evaluation Kits

#R8175

$1,506.00 USD

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IMAP TR-FRET PDE Evaluation Kit (with cAMP and cGMP substrates) - PN: R8176

IMAP TR-FRET PDE Evaluation Kits
Includes optimized protocol, cAMP and cGMP substrates; customer provides their own PDE enzyme

#R8176

$1,506.00 USD

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Resources of Enzyme - IMAP Assays