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.

IMAP Substrates

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

IMAP Assay Kits

Product Name

Description

IMAP Binding Buffers

IMAP Binding Reagent

TR-FRET
Tb-donor

Labeled Substrate

Demo Kit

IMAP® Evaluation Demo Kit

For kinase, phosphatase and Phosphodiesterase fluorescent polarization or TR-FRET assays. Consists of proprietary IMAP Binding Reagent and Binding Buffers, labeld calibrators/substrates, TR-FRET Tb-donor to run a calibration curve for 800 data points in a standard 384 well format.

Evaluation Kits

IMAP® FP Evaluation Kit

For kinase and phosphatase fluorescent polarization assays. Consists of proprietary IMAP Binding Reagent and Binding Buffers, sufficient to generate 800 data points in a standard 384 well format. Labeled substrates are available to purchase separately .

N/A

Order separately

IMAP® TR-FRET Evaluation Kit

For kinase and phosphatase TR-FRET assays. Consists of proprietary IMAP Binding Reagent and Binding Buffers, TR-FRET Tb-donor, sufficient to generate 800 data points in a standard 384 well format. Labeled substrates are available to purchase separately .

Order separately

IMAP® PDE FP Evaluation Kit

For Phosphodiesterase (PDE) fluorescent polarization assays. Consists of proprietary IMAP Binding Reagent and Binding Buffers, labeld cAMP and cGMP substrates, sufficient to generate 800 data points in a standard 384 well format

N/A

IMAP® PDE TR-FRET Evaluation Kit

For Phosphodiesterase (PDE) TR-FRET assays. Consists of proprietary IMAP Binding Reagent and Binding Buffers, labeld cAMP and cGMP substrates, TR-FRET Tb-donor, sufficient to generate 800 data points in a standard 384 well format

Screening Express Kits

IMAP® FP Screening Express Kit

Fluorescence Polarization Screening Express Kit with Progressive Binding System. Consists of proprietary IMAP Binding Reagent and Binding Buffers, sufficient to generate 8,000 data points in a standard 384 well format. Labeled substrates are available to purchase separately .

N/A

Order separately

IMAP® TR-FRET Screening Express Kit

TR-FRET Screening Express Kit with Progressive Binding System. Consists of proprietary IMAP Binding Reagent and Binding Buffers, TR-FRET Tb-donor, sufficient to generate 8,000 data points in a standard 384 well format. Labeled substrates are available to purchase separately .

Order separately

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Ordering Options of Enzyme - IMAP Assays

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imap-assay-kit, imap-assay-components

#R8166

#R8155

#R8161

#R8175

#R8176

#R8127

#R8160

#R7470

Tb-donor for IMAP TR-FRET assay, sufficient amount for screening and generate 8,000 data points in a standard 384 well format.

#R7471

Tb-donor for IMAP TR-FRET assay, sufficient amount for high throughput screening and generate 50,000 data points in a standard 384 well format.

#R7505

Fluorescence labeled cAMP substrate 20 nmoles for IMAP phosphodiesterase (PDE) assays, sufficient for 8,000 data points based on 100 nM in reaction. Replaced R7091

#R7506

Fluorescence labeled cAMP substrate 120 nmoles for IMAP phosphodiesterase (PDE) assays, sufficient for 50,000 data points based on 100 nM in reaction.

#R7507

Fluorescence labeled cGMP substrate 20 nmoles for IMAP phosphodiesterase (PDE) assays, sufficient for 8,000 data points based on 100 nM in reaction. Replaced R7090

#R7508

Fluorescence labeled cGMP substrate 120 nmoles for IMAP phosphodiesterase (PDE) assays, sufficient for 50,000 data points based on 100 nM in reaction.

#R7110

Fluorescence labeled peptide 5FAM-GRPRTSSFAEG-COOH, substrate for Akt1, Akt2, Akt3, MSK1, MSK2, SGK1, Plk3, sufficient for 8,000 data points based on 100 nM in reaction

#R7129

Fluorescence labeled peptide LVEPLTPSGEAPNQK-5FAM-COOH, substrate for JNK1, JNK2,JNK3, p38 (α,β,γ,δ isoforms), sufficient for 8,000 data points based on 100 nM in reaction

#R7157

Fluorescence labeled peptide 5FAM-KVEKIGEGTYGVV-NH2, substrate for Src, Fyn, Lck, Yes, Hck, Rse, sufficient for 8,000 data points based on 100 nM in reaction

#R7172

Fluorescence labeled peptide 5FAM-KVEKIGEGTYGVV-NH2, substrate for Src, Fyn, Lck, Yes, Hck, Rse, sufficient for 50,000 data points based on 100 nM in reaction

#R7184

Fluorescence labeled peptide 5FAM-AKRRRLSSLRA-COOH, substrate for ROCK-II,Rsk1,Rsk2,Rsk3,Tab1, sufficient for 8,000 data points based on 100 nM in reaction

#R7229

Fluorescence labeled peptide 5FAM-AKRRRLSSLRA-COOH, substrate for ROCK-II,Rsk1,Rsk2,Rsk3,Tab1, sufficient for 50,000 data points based on 100 nM in reaction

#R7250

Fluorescence labeled peptide 5FAM-GRTGRRNSI-NH2, substrate for Aurora A, PKA, PKG, sufficient for 8,000 data points based on 100 nM in reaction

#R7252

Fluorescence labeled peptide 5FAM-GGGPATPKKAKKL-COOH, substrate for CDK1/Cyclin B, CDK2/Cyclin A, CDK2/CyclinE, CDK3/CyclinE, CDK5/p25, CDK5/p35,CDK6/Cyclin D3, sufficient for 8,000 data points based on 100 nM in reaction

#R7254

Fluorescence labeled peptide 5FAM-GRHDSGLDSMK-NH2, substrate for IKKβ, IKKα, sufficient for 8,000 data points based on 100 nM in reaction

#R7255

Fluorescence labeled peptide 5FAM-GRTGRRNSI-NH2, substrate for Aurora A, PKA, PKG, sufficient for 50,000 data points based on 100 nM in reaction

#R7257

#R7292

Fluorescence labeled peptide IPTTPITTTYFFFK-5FAM-COOH, substrate for Erk1, Erk2, p38 (α,β,γ,δ isoforms), sufficient for 8,000 data points based on 100 nM in reaction

#R7307

Fluorescence labeled peptide LVEPLTPSGEAPNQK-5TAMRA-NH2, substrate for JNK1, JNK2, JNK3, p38 (α,β,γ,δ isoforms), sufficient for 8,000 data points based on 100 nM in reaction

#R7311

Fluorescence labeled peptide 5FAM-HAAIGDDDDAYSITA-NH2, substrate for CK1, CK1δ, sufficient for 8,000 data points based on 100 nM in reaction

#R7312

Fluorescence labeled peptide 5FAM-HAAIGDDDDAYSITA-NH2, substrate for CK1, CK1δ, sufficient for 50,000 data points based on 100 nM in reaction

#R7313

Fluorescence labeled peptide 5TAMRA-GRTGRRNSI-COOH, substrate for PKA, PKG, sufficient for 8,000 data points based on 100 nM in reaction

#R7319

Fluorescence labeled phosphopeptide LVEPL-pT-PSGEAPNQ(K-5FAM)-COOH, substrate for assay calibration, sufficient for 8,000 data points based on 100 nM in reaction

#R7352

Fluorescence labeled peptide 5TAMRA-YSPTSPSYSPTSPSYSPTSPS-COOH, substrate for CDK7, sufficient for 8,000 data points based on 100 nM in reaction

#R7383

Fluorescence labeled peptide 5TAMRA-KKKSPGEYVNIEFG-NH2, substrate for Ros, Met, sufficient for 50,000 data points based on 100 nM in reaction

#R7434

Fluorescence labeled peptide 5FAM-IPTTPITTTYFFFK-NH2, substrate for Erk1, Erk2, p38 (α,β,γ,δ isoforms), sufficient for 8,000 data points based on 100 nM in reaction

#RP7005

Fluorescence labeled peptide 5FAM-LKKLRRRSDANF-NH2, substrate for CaMKI, sufficient for 8,000 data points based on 100 nM in reaction

#RP7030

Fluorescence labeled peptide 5FAM-RKRQGSVRRRVH-OH, substrate for PKC, sufficient for 8,000 data points based on 100 nM in reaction

#RP7032

Fluorescence labeled peptide 5FAM-RFARKGSLRQKNV-COOH, substrate for PKCζ, sufficient for 8,000 data points based on 100 nM in reaction

#RP7035

Fluorescence labeled peptide 5FAM-GRRESLTSFG-NH2, substrate for PKA, PKG, sufficient for 8,000 data points based on 100 nM in reaction

#RP7045

Fluorescence labeled peptide 5FAM-PLSRTLSVSSLPGL-NH2, substrate for c-TAK, sufficient for 8,000 data points based on 100 nM in reaction

#RP7046

Fluorescence labeled peptide 5FAM-GEILSRRPSYRK-NH2, substrate for MSK1, sufficient for 8,000 data points based on 100 nM in reaction

#RP7116

Fluorescence labeled peptide 5FAM-YSPTSPSYSPTSPSYSPTSPS-OH, substrate for TKL, STE, CK1, CMGC, sufficient for 8,000 data points based on 100 nM in reaction

#RP7119

Fluorescence labeled peptide 5FAM-EPPQSQEAFADLWK-NH2, substrate for TKL, STE, CK1, CMGC, sufficient for 8,000 data points based on 100 nM in reaction

#RP7120

Fluorescence labeled peptide 5FAM-RRRFRPASPLRGPPK-OH, substrate for Dyrk1A, TKL, STE, CK1, CMGC, sufficient for 8,000 data points based on 100 nM in reaction

#RP7123

Fluorescence labeled peptide 5FAM-ATGPLSPGPFGRR-OH, substrate for Erk2, TKL, STE, CK1, CMGC, sufficient for 8,000 data points based on 100 nM in reaction

#RP7131

Fluorescence labeled peptide 5FAM-FLTEYVATRWYRAPEIMLN-NH2, substrate for MAPK, TKL, STE, CK1, CMGC, sufficient for 8,000 data points based on 100 nM in reaction

#RP7140

Fluorescence labeled peptide 5FAM-GTFRSSIRRLSTRRR-OH, substrate for Nek2, IRAK4, sufficient for 8,000 data points based on 100 nM in reaction

#RP7153

Fluorescence labeled peptide 5FAM-GRSRSRSRSR-OH, substrate for SPRK1, IRAK4, sufficient for 8,000 data points based on 100 nM in reaction

#RP7548

Fluorescence labeled peptide 5FAM-ERMRPRKRQGSVRRRV-NH2, substrate for PKCγ, Pim1, sufficient for 50,000 data points based on 100 nM in reaction

#RP7609

Fluorescence labeled peptide 5FAM-KKRKSSLRRWSPLTPRQMSFDC-NH2, substrate for PKA, PKC, CAMK, MAPK, TKL, STE, CK1, CMGC, sufficient for 50,000 data points based on 100 nM in reaction

#RP7616

Fluorescence labeled peptide 5FAM-YSPTSPSYSPTSPSYSPTSPS-OH, substrate for TKL, STE, CK1, CMGC, sufficient for 50,000 data points based on 100 nM in reaction

#RP7640

Fluorescence labeled peptide 5FAM-GTFRSSIRRLSTRRR-OH, substrate for Nek2, IRAK4, sufficient for 50,000 data points based on 100 nM in reaction

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