HTRF KINEASE STK S3 KIT - 20K PTS 丝氨酸激酶测量试剂盒S3-20°C,000测试

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 protocol

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 Eu-cryptate and XL665 conjugates are added. The detection buffer contains EDTA to stop the enzymatic reaction.