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Enzymatic Assay

As the simplest biological reaction system, enzymatic assays can accurately reflect the ability of the compounds to be evaluated to inhibit or activate specific enzymes. Although the enzymatic reaction system is simple, developing an enzymatic system that can be used to evaluate compounds is still challenging.  

For the development and verification of an enzymatic experiment, the following steps are usually required: 

    Confirm the appropriate substrate and determine The range of linear relationship between reaction signal and enzyme concentration; 

    Select the appropriate reaction volume (microplate) and optimize the reaction buffer; 

    Measurement of Michaelis-Menten constant to determine ATP concentration; 

    Measurement of reaction kinetics; 

    Use control compounds for method verification; 

    The method will be further optimized and verified during the project.  

At Truwaybio, we will optimize each enzymology method according to the above steps.  

The throughput of enzymatic experiments can be screened in 96-well plates or 384-well plates.  

Reaction types include TR-FRET/HTFR, Alpha-screen, AlphaLISA, Transcreener assay, ADP-Glo™ Kinase Assay etc.  

The enzymatic methods that Truwaybio has developed and verified include: TRKA wt, TRKA G595R, TRKA G667C, TRKB wt, TRKC wt, TRKC G696A, TRKC G623C, ALK, MET, MET F1200I, ROS1, etc..

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