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ProFluor® PKA Assay

This product is discontinued

We are sorry, but this product is discontinued.

For a compatible replacement, contact Technical Service for assistance.

Technical Service

ProFluor® PKA Assay

"Add-Mix-Read" Convenience for High-Throughput PKA Screening

The ProFluor® PKA Assay measures protein kinase A activity using purified kinase in a multiwell plate format and involves "add-mix-read" steps only—ideal for high-throughput applications.

The assay begins with a standard kinase reaction performed with a provided Bisamide Rhodamine 110 peptide substrate (PKA R110 Substrate). In this configuration the substrate is nonfluorescent. Following the kinase reaction, a termination buffer containing a Protease Reagent is added, simultaneously stopping the kinase reaction,  removing amino acids specifically from the nonphosphorylated PKA R110 Substrate, and producing highly fluorescent Rhodamine 110. The phosphorylated PKA R110 Substrate is resistant to digestion by the Protease Reagent and remains nonfluorescent. Thus, the fluorescence intensity measured in the assay is inversely correlated with kinase activity.

Minimal Test Compound Interference

The Rhodamine 110 fluorescent signal produced is much higher than the fluorescent signal given off by test compounds. The assay produces excellent Z´-factor values (>0.8) in either 96- or 384-well plate formats and easily distinguishes known PKA inhibitors from other compounds.

Learn more about our custom options for this product at: www.promega.com/custom/

Protocols

Specifications

What's in the box?

Item Part # SizeConcentrationAvailable Separately

rATP

P113C 1 x 250μl10mM View Product

PKA R110 Substrate

V304A 1 x 12μl

Stabilizer Reagent

V306A 1 x 12μl

5X Reaction Buffer A

V307A 1 x 6ml

5X Termination Buffer A

V308A 1 x 5ml

Protease Reagent

V309A 1 x 240μl

SDS

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Certificate of Analysis

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Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

BB

Store at –20°C.

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