Journal: Journal of Biological Chemistry

Article Title: The Human Mitochondrial Transcription Termination Factor (mTERF) Is FullyActive in Vitro in the Non-phosphorylatedForm

doi: 10.1074/jbc.m501145200

Figure Lengend Snippet: FIG. 3. The active mTERF protein is not phosphorylated. A, phosphoprotein blot staining was performed as described under “Ma- terials and Methods.” Ovalbumin (OA) has two phosphorylated amino acid residues and was used as a positive control. Bovine serum albumin (BSA) is not phosphorylated and was used as a negative control. MW, Precision Plus protein standards (Bio-Rad). B, two- dimensional gel electrophoresis analysis of recombinant mTERF. Pu- rified mTERF protein (fraction 69; see Fig. 2B) was separated by isoelectric focusing (pH 7–11) in the first dimension and by electro- phoresis with a 12.5% SDS-polyacrylamide gel in the second dimen- sion. Proteins were visualized by Coomassie Blue staining. MW, Precision Plus protein standard plug (Bio-Rad). C, two-dimensional gel electrophoresis was performed as for panel B. Samples were incubated at 37 °C for 30 min in the absence (top) or presence (bottom) of 4 units of alkaline phosphatase (AP) as described under “Materials and Methods.” Proteins were visualized by silver staining. 1st dim, first dimension.

Article Snippet: Precision Plus protein standard plug (BioRad) was used as a molecular mass marker.

Techniques: Staining, Positive Control, Negative Control, Two-Dimensional Gel Electrophoresis, Electrophoresis, Recombinant, Incubation, Silver Staining