Figure 3
From: Binding of the baculovirus very late expression factor 1 (VLF-1) to different DNA structures
The effect of monoclonal antibody HA.11 and glutaraldehyde fixation on electrophoretic mobility of *VLF-1 complexes with cruciform DNA. Reaction mixtures (10 μl) containing 0.004 pmol 32P-labeled cruciform DNA (substrate V) and 40 ng of *VLF-1 were preincubated for 30 min at 23°C. No *VLF-1 was added to the reactions shown in lanes 1, 7, and 13. After preincubation, the monoclonal antibody HA.11 (α-HA) was added at the following final dilutions: lanes 3 and 9, 1:6,000; lanes 4 and 10, 1:2,000; lanes 5 and 11, 1:600; and lanes 6, 12, and 13, 1:200, and the incubation was continued for 10 min. The reactions shown in lanes 7–13 were fixed with 0.5% glutaraldehyde prior electrophoresis, whereas those shown in lanes 1 to 6 were analyzed by electrophoresis in a native 6% polyacrylamide gel without fixation with glutaraldehyde. The position of the *VLF-1 complexes with cruciform DNA is indicated by an asterisk. The supershifted bands formed in the presence of monoclonal antibody HA.11 are indicated by double asterisks.