Assay Method Information

Assay Name:  Assay of Inhibition of Staphylococcus Aureus RNA Polymerase
Description:  Fluorescence-detected RNA polymerase assays with S. aureus RNA polymerase were performed as in Example 21.1, using reaction mixtures containing (20 μl): 0-100 nM test compound, 75 nM S. aureus RNA polymerase core enzyme, 300 nM S. aureus σ A, 20 nM 384 bp DNA fragment containing the bacteriophage T4 N25 promoter, 100 μM ATP, 100 μM GTP, 100 μM UTP, 100 μM CTP, 40 mM Tris-HCl, pH 8.0, 80 mM NaCl, 5 mM MgCl2, 2.5 mM DTT, and 12.7% glycerol. IC50 is defined as the concentration of inhibitor resulting in 50% inhibition of RNA polymerase activity.
Affinity data for this assay
 

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