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In general terms, a laboratory or company interested in high-throughput screening for drug discovery will have already established an assay for the enzyme or system of interest to them. This might be, for example, an enzyme with an activity associated with a particular disease state, or a receptor they believe to be involved in particular signaling cascade, or a newly discovered enzyme that they wish to characterize, etc. There is essentially no limit to the number of topics that can be addressed this way. What is important is that the customer is interested in screening broad populations of compounds at the same time. Very often this process is automated robotically to allow for faster processing without the risk of human error. It is completely up to the customer to develop/test/troubleshoot the assay for their system of interest. Once the assay system has been established, the screening process can actually begin. Multiple reaction wells will be set-up (often in a 96-well plate format, although 384-well plates and other formats can certainly be used as well). An individual compound from the library is then added to each reaction well.Depending on the assay type, the read-out is further assessed by comparing treated with controls. Compounds that either increase or decrease the signal (depending upon the experiment in question) would be considered “hits” or “lead compounds / lead candidates”. Following initial screening, lead candidates are further evaluated with additional assays. This could involve assays with purified enzymes to determine inhibitor constants, kinetic studies, etc.