Enzyme - IMAP Assays
IMAP® technology provides a homogeneous assay applicable to a wide variety of kinases, phosphatases, and phosphodiesterases without regard for substrate peptide sequence. The assay is a simple mix-and-read procedure allowing accurate determination of enzyme activity.
- IMAP provides a complete assay system for screening kinases, phosphatases, and phosphodiesterases
- Because IMAP assays are not antibody-based, they are generic and can be used for any kinase, phosphatase, or phosphodiesterase
- Robust fluorescence signal gives reliable results with good Z factors
- IMAP assays are homogeneous and amenable to miniaturization for greater cost savings
- IMAP assays are available in both FP and TR-FRET detection modes to meet users' screening needs
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 Progressive Binding System lets researchers optimize their FP and TR-FRET assay conditions for each substrate used. The system consists of Progressive Binding Buffer A and Progressive Binding Buffer B, along with Progressive Binding Reagent. The two buffers and the reagent can be combined in different proportions according to the acidic character of the substrate choice, desirable ATP concentrations and background parameters.
Molecular Devices offers a wide range of validated substrates and calibrators. Substrates may be used with the enzymes for which they were originally validated, or as potential substrates for other enzymes.
• Validated IMAP substrates with optimized binding conditions ensure the best performance when using the IMAP platform
• Substrates come in two sizes and can be used with both IMAP FP and IMAP TR-FRET
• Dozens of different substrates have been validated with over 100 enzymes. Many substrates are available with either red or green fluorescent label, enabling multiplexing and providing a way to address issues arising from compound autofluorescence
Data of IMAP Assays
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.
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
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
IMAP FR PDE Evaluation Kit - PN: R8175
IMAP FP PDE Evaluation Kits
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
Resources of Enzyme - IMAP Assays
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…
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…
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.
Get information about proprietary IMAP Technology which provides a non-radioactive, homogeneous assay for kinase activity.
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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
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…