One of the best ways to measure kinase activity in a high throughput format is through the use of a biochemical kinase assay kit. Kinase enzymes are capable of transferring phosphate groups from ATP to a substrate that leaves ADP and phosphorylated molecule. When it comes to a universal kinase activity assay, the top platform is Transcreener. It uses a highly specific antibody that relies ADP detection to properly measure the kinetics of an enzyme.
Why Direction Detection
The reason that direction detection is the best option is because it eliminates the coupling steps that are known to be complex. This means that there is less time spent on addition steps during screens and lowers the possibility of interference to compounds. This detection method is available in three fluorescent methods that will likely fit the hardware you are already have in-house. You can simply pick which option works best for you and use it for all your drug discovery needs.
Available Detection Formats
Depending on the type of fluorescent plate reader that you already own, you can select the corresponding detection method. Your options consist of the following:
TR-FRET – Time-Resolved Forster-Resonance-Energy-Transfer
FP – Fluorescent Polarization
FI – Fluorescent Intensity
Use With Substrates
While these are easy to use with a one step mix-and-read procedure, they can not only be used with variety kinase enzymes but also virtually any substrate. Carbohydrate kinases, protein, kinases, and lipid kinases are all capable of being used with this simple procedure.
Being used for various substrates is only one of the benefits of this option. It can be used in multiple different applications in your research and development. The following are some of the most common:
* Ability to determine the activity of enzymes
* Ablity to screen for small molecule inhibitors and activators
* May be used for selectivity profiling
* Can determine accurate IC50s
* Able to measure lead molecule residence times
At Bellbrook Labs, we offer kinase activity assays as well as a variety of resources related to the topic. Our kinase assay kits are competitive immunoassays and are implemented in a simple mix-and-read format. In addition, Bellbrook Labs provides other products and services related to the drug discovery process. You can learn more or order our products at www.BellbrookLabs.com.
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