One method for detection and quantification of cell proliferation is based on the incorporation of nucleoside analogs like [3H]thymidine or 5-bromo-2’-deoxyuridine (BrdU) to into the cells’ DNA during replication and visualization by autoradiography or with an anti-BrdU-antibody respectively. Both methods exhibit several limitations. Working with [3H]thymidine is troublesome because of its radioactivity and autoradiography is slow and thus not suitable for rapid high-throughput studies. The major disadvantage of BrdU staining is that the double-stranded DNA blocks the access of the anti-BrdU antibody to BrdU units. Therefore, samples have to be subjected to harsh denaturing conditions resulting in degradation of the structure of the specimen.
The PromoKine EdU-based Cell Proliferation Kits overcome these limitations, providing a superior alternative to BrdU and [3H]thymidine assays for directly measuring DNA synthesis flow cytometry.
EdU (5-ethynyl-2’-deoxyuridine) is a nucleoside analog to thymidine and is incorporated into DNA during active DNA synthesis. In contrast to BrdU assays, the EdU-based Cell Proliferation Assays are not antibody based and therefore do not require harsh DNA denaturation for detection of the incorporated nucleoside. Instead, the EdU-based Cell Proliferation Assays utilize click chemistry for easily coupling fluorophores to the incorporated EdU – allowing fast and sensitive detection in a variety of dye fluorescent readouts. Furthermore, the streamlined detection protocol reduces the total number of steps and significantly decreases the total amount of time. The simple click chemistry detection procedure is completed within 30 minutes and is compatible with multiplexing for content and context-rich results.