Fast chemiluminescent measurement of RNA polymerase activity based on photon counting technology.
A fast and simple assay for T7 RNA polymerase based upon chemiluminescent detection of the synthesized, digoxigenin-labeled RNA on a nylon membrane with anti-digoxigenin coupled alkaline phosphatase and CSPD as substrate is described. Activity of RNA polymerase is determined with high sensitivity by quantifying the emitted light of the microplate-formatted dot-blot membrane with a photon counting microplate luminometer and a specially designed filter adapter. The described method is one example for the application of this new adapter to measure luminescent membrane filters.
Walter
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N
Abteilung Biochemische Kinetik, Max-Planck-Institut für Biophysikalische Chemie, Göttingen, FRG.
Steiner
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C
D012321:DNA-Directed RNA Polymerases / Q000032:analysis* / Q000378:metabolism; D004076:Digoxigenin; D005374:Filtration / Q000295:instrumentation; D007124:Immunoenzyme Techniques; D008163:Luminescent Measurements; D008567:Membranes, Artificial; D009757:Nylons; D010783:Photometry / Q000379:methods*; D017785:Photons; D012313:RNA / Q000032:analysis / Q000378:metabolism; D014764:Viral Proteins
D008567:Membranes, Artificial; D009757:Nylons; D014764:Viral Proteins; D012313:RNA; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases; D004076:Digoxigenin