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. 1998 Jun 15;12(12):1884–1893. doi: 10.1101/gad.12.12.1884

Figure 2.

Figure 2

 Characterization of the holoenzyme and activator preparations and transcription inhibition by monoclonal antibody 2G10. (A) SDS-PAGE of the E. coli RNAP (lanes 1,2); RcRNAP (lanes 3,4); RcNtrCC3 (lane 5) and RcNtrCC7 (lane 6); standards with sizes indicated in kD. (B) Western analysis of the RcRNAP and EcRNAP purification by use of mAb 2G10, specific for E. coli σ70. Fractions are from E. coli and R. capsulatus as follows: (Lanes 1,5) PEG precipitation fraction; (lanes 2,6) heparin agarose fraction; (lanes 3,7) DEAE–Sepharose fraction; (lane 4) HPLC-purified EcRNAP/σ70 holoenzyme. (C) RcNtrC activated transcription is abolished by σ70 mAb 2G10. The nifA1Mut1 promoter was used. (Lane 1) no activator; (lanes 2–5) 350 nm RcNtrCC7; (lanes 3–5) 270 nm MBP–NtrB; (lane 4) 1 μl of mAb 2G10; (lane 5) 1 μl of control mAb.