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HTRF KinEASE STK Discovery Detection Kit, 1,000 Assay Points
The HTRF KinEASE assay series is a semi-universal biochemical platform to study kinase activity. The Discovery kit provides a straightforward way to determine which of the STK kits must be used for a given serine/threonine kinase.
For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption & disposal requirements under European REACH regulations (EC 1907/2006).
Product information
Overview
The HTRF KinEASE kit series provides a simple biochemical approach to study kinase activity, screen for inhibitors and characterize them. Three STK and one TK kits, offer a semi-universal method for assessing phosphorylation on Serine/Threonine (STK) and Tyrosine (TK) residues, respectively.
Over 188 kinases have been validated with this technique and the assay validation section of this page includes technical tips for each assay setup.
The Discovery kit is intended to help determine which of the three STK kits is appropriate when working with a non-validated Serine/Threonine kinase.
Specifications
| Assay Points |
1000
|
|---|---|
| Assay Target Type |
Kit
|
| Assay Technology |
HTRF
|
| Brand |
HTRF
|
| Quantity |
1
|
| Shipping Conditions |
Shipped Ambient
|
| Therapeutic Area |
Metabolism/Diabetes
NASH/Fibrosis
Oncology & Inflammation
|
| Unit Size |
1,000 Assay Points
|
Video gallery
How it works
Assay principle
Each HTRF KinEASE kit is based on the same assay format, and uses a universal biotinylated substrate, a phospho-specific Eu-cryptate-labeled monoclonal antibody, and SA-XL665. All three STK peptidic substrates are made of the same sequence for antibody detection, next to a variable sequence recognized by enzymes. The targeted residue for phosphorylation lies in between as part of both sequences for enzymatic activity (unphosphorylated) and antibody detection (phosphorylated). It has been shown that 3 different variable sequences could enable the study of most Serine/Threonine kinases.
Assay principle
As shown here, all HTRF KinEASE assays work the same way and involve two steps: Step 1: Kinase reaction The kinase is incubated in the presence or absence of compounds and the appropriate substrate. The supplemented enzymatic buffer is added in the kinase buffer for tyr kinases only. ATP is added to start the reaction. Step 2: Detection Eu3+ cryptate and XL665 conjugates are added. The detection buffer contains EDTA to stop the enzymatic reaction.
Assay validation
Validated kinases
Click in the table below to view the list of validated enzymes, the kit which use is recommended, and download the technical datasheet including experimental setting and validation data. Non listed STKs can be assessed with the Discovery kit to find out which HTRF KinEASE STK kit may be used.
Over 190 Serine/Threonine and Tyrosine kinases have already been validated with the HTRF KinEASE kits, and their optimal substrate determined. Note that one given enzyme may be compatible with several substrates, as shown here with MST1 that work equally well with S3 and S2, but significantly less with S1.
Resources
Guide
HTRF solutions, guide to major applications
This guide provides you an overview of HTRF applications in several therapeutic areas.
Guide
The ultimate guide to successful kinase binding experiments
Your guide for mastering the art of kinase binding assay optimizations
This complete guide provides you with all the tools you need...
SDS, COAs, Manuals and more
Are you looking for technical documents for this product. We have housed them in a dedicated section., click on the links below to explore.
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