Accelerating the pace of luciferase reporter gene assays.
Luciferase reporter gene assays have gained more importance because of their easy readout, high sensitivity and lack of environmental waste disposal problems. However, several obstacles remain that have prohibited a wider use and the implementation of this type of assay in high-throughput screening programs: (i) Measurements need to be carried out within an active enzyme reaction, and the assessment of such reactions are time-dependent; (ii) the signal produced has a "flash" type characteristic and therefore requires specialized equipment for measurement; and (iii) side-reactions can occur that interact with the signal readout of the assay in a non-reproducible way. These hurdles make an otherwise convenient assay principle troublesome for larger-scale screening use. We have attempted to overcome these problems by different means, leading to the development of LucLite, a stable signal homogeneous reagent system. This system allows use in a higher throughput screening capacity and enables the use of standard scintillation/luminescence instruments.
Roelant
C H
CH
Packard Instrument B. V., Groningen, The Netherlands.
Burns
D A
DA
Scheirer
W
W
D001681:Biological Assay; D002460:Cell Line; D017930:Genes, Reporter / Q000235:genetics*; D005820:Genetic Testing / Q000379:methods*; D006801:Humans; D007202:Indicators and Reagents; D008156:Luciferases / Q000235:genetics*; D008163:Luminescent Measurements; D015398:Signal Transduction / Q000502:physiology; D013601:T-Lymphocytes / Q000166:cytology; D014162:Transfection
D007202:Indicators and Reagents; D008156:Luciferases