Chemiluminescence-based BrdU ELISA to measure DNA synthesis.
We describe a simple, sensitive, nonradioactive, relatively rapid and relatively inexpensive protocol to measure DNA synthesis in cultured cells by a chemiluminescent bromodeoxyuridine (BrdU) enzyme-linked immunosorbent assay (ELISA). We show that it exhibits similar sensitivity and activity as traditional 3H-thymidine incorporation assays and a commercial chemiluminescent BrdU ELISA kit when tested in commonly used cell lines, such as NIH 3T3 cells, mink lung epithelial cells (Mv1LU), and baby hamster kidney (BHK-21) fibroblasts. This assay also exhibits a wider dynamic range than colorimetric BrdU ELISA methods. Besides being a viable, nonradioactive alternative to 3H-thymidine incorporation assays, our BrdU ELISA is less expensive than a commercial chemiluminescent BrdU ELISA kit.
Hawker
James R
JR
Cardiovascular Research Institute and Department of Medical Physiology, College of Medicine, Texas A&M University System Health Science Center, Medical Research Building, Rm 206A, 702 SW H. K. Dodgen Loop, Temple, TX 76504, USA.
D016475:3T3 Cells; D000818:Animals; D001973:Bromodeoxyuridine / Q000032:analysis*; D002455:Cell Division; D002460:Cell Line; D003124:Colorimetry; D006224:Cricetinae; D004247:DNA / Q000096:biosynthesis*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D008163:Luminescent Measurements; D051379:Mice; D013936:Thymidine / Q000032:analysis
D004247:DNA; D001973:Bromodeoxyuridine; D013936:Thymidine