Our revolutionary assay technology leverages immobilized kinase substrates on a 96-well plate for precise analysis. With the ability to accommodate over 100 different types of active kinases, this innovative approach facilitates the phosphorylation of substrate mixtures on the plate. The resulting phosphorylation events can be accurately detected and quantified using our highly specific phospho-specific antibodies. Notably, these assay kits offers the convenience of bench-top performance, eliminating the need for radioactivity while providing a colorimetric readout. Enjoy the benefits of a non-radioactive and colorimetric solution in our rapid assay kits for your kinase analysis needs.

This kit is designed for testing the activity of a variety of serine/threonine kinases (STK) and screening the inhibitors/activators of corresponding kinases. The principle of the assay, illustrated in Figure 1., is:

1. The kinase substrates are immobilized onto the 96-well plate.
2. The kinases use ATP to phosphorylate the substrates on the plate.
3. The phosphorylated product is detected by the phospho-substrate specific antibodies.

Figure 1. Kinase assay kit principle.

 

The Type I Kinase Assay Kits are designed for the assay of PKA in solution. Purified or partially purified PKA will phosphorylate the PKA substrate on the 96-well plate with the presence of ATP. The phosphorylated substrate (pSubstrate) is then analyzed with the anti-phosphosubstrate antibodies. Subsequently, the binding anti-pSub antibodies will be quantified with the secondary antibody-HRP conjugate, which generates the color signal from its substrate, TMB (OD at 450 nm). The final color intensity is proportional to the initial PKA phosphorylation activity.

 

Assay for activity of kinase using Type II PKA kinase assay kit (ICP0217).
(A: 100 ng; B: 50 ng; C: 25 ng; D: 12 ng; E: 6 ng; F: 0.00 ng)

The Type II PKA Kinase Assay Kits are designed for kinase solid affinity matrices such as IP kinases or PKAs on GSH beads. They are also useful for analyzing activity of PKA in the solution phase. The principle of the assay is that the PKA kinase phosphorylates the substrate-BSA conjugates (component C) in the presence of ATP. The phosphorylated BSA-substrate is then analyzed using SDS-PAGE resolution and immunoblotting techniques.