When it comes to the significant process that is compiling components for the correct kinase assay for your work, it can be extremely time-consuming and expensive. Having a simple mix-and-read method can take a lot of the stress out of this process. The good news is that you can find a kinase assay kit which does exactly what you need and is compatible with the plate reader you already have on hand.
Applications of Kinase Activity Assays
When you have the right product in hand, it can remove the need for time-consuming assay development. Universal activity assays can drastically reduce development time within a family of enzymes by detecting the invariant enzyme product, which in this case is ADP. From these products you can expect the following:
The potential to screen for small molecular inhibitors
The ability for inhibitor selectivity screening
A universal method to determine enzymatic activity
Potential for measuring the residence time/off-rate-/ dissociation of lead molecules and the target
Allowing mechanism of action studies
Ability to determine IC50 values to measure inhibitor potency
How It Works
With universal kinase assay, there is a requirement to directly detect ADP. This is done using a specific antibody which binds specifically to ADP. When kinase assays are run under initial velocity conditions, the selectivity of the antibody allows direct detection of ADP even in the presence of large amounts of ATP. The ADP antibody allows for sensitively to detecting ADP all the way down to 10 nM with a Z’ value of 0.7 or higher.
Continuous or Endpoint Mode Formats
These kinase activity assays, provide a mix-and-read format that is homogenous, simplifying automation while ensuring the highest level of flexibility in terms of assay protocol. Continuous assays are typically used during assay development and for residence time determination, while endpoint assays are more common for HTS.
Flexibility in Terms of Plate Reader
There are three different detection modes available and working with common plate readers is possible. It’s crucial to look over the technical manual for your assay to see what the best instrument settings and filter sets are. Be sure your instrument is a fit with the assays available. If you’re interested in learning more, Bellbrook Labs can help. You can reach us at www.bellbrooklabs.com.