PKA Assay Kit

A sensitive, non-radioactive, homogeneous and rapid assay for PKA

• Uses fluorescence polarization detection methods
• Assays PKA serine/threonine kinase
• Available in kit and bulk packaging
• Includes enzyme obtained from Upstate

Non-radioactive screening of PKA activity is now possible using fluorescence polarization assays. The Molecular Devices IMAP FP PKA Assay Kit provides a sensitive and rapid assay that detects the activity of PKA without antibodies in a homogeneous format. For convenience and economy, the reagents are available as a kit for assay evaluation and in bulk for high-throughput screening.

Intended Use

This assay kit is designed for use in drug discovery to screen the activity of PKA in the presence of putative drug compounds. It also enables the user to validate the IMAP Platform in a well-defined system.

Principle of the Assay

The IMAP® technology is based on the high affinity binding of phosphate at high salt concentration by immobilized metal (MIII) coordination complexes on nanoparticles. This IMAP “binding reagent” complexes with phosphate groups on phosphopeptides generated in a kinase reaction. For FP detection such binding causes a change in the rate of the molecular motion of the peptide, and results in an increase in the fluorescence polarization value observed for the fluorescein label attached at the end of the peptide (Figure 1). This assay, unlike antibody-based kinase assays, is applicable to a wide variety of kinases without regard to the substrate peptide sequences.

		Figure 1. Diagram of IMAP assay principle.

Kit Contents

The kit contains a fluorescent Substrate, IMAP Binding Reagent, IMAP Binding Buffer, Kinase Reaction Buffer, and PKA enzyme (Upstate). When used as suggested, each kit provides sufficient reagents for 800 assays in 384-well plate format. Bulk kits contain reagents for 8,000 assay points in 384-well plate format.

Assay Conditions

The fluorescence polarization value of the sample is measured with any instrument in the Molecular Devices Analyst® line or SpectraMax M5.

		Figure 2. Diagram of IMAP assay principle.

Applications

Measure the kinase activity of PKA.

Assay Performance

		Figure 3. Representative titration of PKA with the IMAP FP PKA Assay Kit (SoftMax® Pro graph).

Ordering Information

Product Number	R8110	R8117
Product Name	IMAP® FP PKA Kinase Explorer Assay Kit	IMAP® FP PKA Kinase Bulk Assay Kit
Unit Size	1 Kit provides reagents sufficient for 800, 20 µL reactions	1 Bulk Kit provides reagents sufficient for 8000, 20 µL reactions
Components	IMAP Progressive Binding Reagent (400X) IMAP Progressive Binding Buffers Kinase Reaction Buffer incl. 0.1% BSA PKA, active Substrate (5-carboxy- fluorescein-labeled)	IMAP Progressive Binding Reagent (400X) IMAP Progressive Binding Buffers Kinase Reaction Buffer incl. 0.1% BSA PKA, active Substrate (5-carboxy- fluorescein-labeled)

To place an order, call your local Molecular Devices Office or Molecular Devices Distributor.