Journal: Molecular microbiology

Article Title: The role of the G2 box, a conserved motif in the histidine kinase superfamily, in modulating the function of EnvZ.

doi: 10.1046/j.1365-2958.2002.03061.x

Figure Lengend Snippet: Fig. 1. Binding interaction between His10-OmpR and various EnvZc G2 box mutants. Aliquots of 4 mg of various EnvZc proteins (EnvZc, EnvZc G401A, G403A and G405A mutants) were each incubated in binding buffer with or without 5 mg of His10-OmpR in a final volume of 100 ml for 45 min at room temperature. Then, pre-equilibrated (with binding buffer) Ni-NTA resin (5 ml) was added into the mixtures, and the binding mixtures were incubated further at 4∞C for 45 min with gentle shaking. After washing three times with binding buffer containing 350 mM NaCl, resin-bound proteins were eluted with binding buffer containing 100 mM EDTA. Eluates were then subjected to SDS–PAGE and Coomassie brilliant blue staining.

Article Snippet: The gel was stained with Coomassie brilliant blue, and the amounts of proteins were estimated using a Bio-Rad Model GS-670 imaging densitometer.

Techniques: Binding Assay, Incubation, Gentle, SDS Page, Staining