Journal: The FEBS journal

Article Title: Purification, microsequencing and cloning of spinach ATP-dependent phosphofructokinase link sequence and function for the plant enzyme.

doi: 10.1111/j.1742-4658.2006.05590.x

Figure Lengend Snippet: Fig. 1. Silver-stained 12% SDS ⁄ PAGE of spinach PFK from dif- ferent purification steps. Lane 1: crude extract. Lane 2: DEAE fractogel eluate. Lane 3: Sephacryl S-400 HR eluate. Lane 4: con- centrated protein after sucrose gradient centrifugation. Lane 5: reactive red eluate. Lane 6: Mono Q eluate. Lane 7: hydroxylapatite eluate. M: molecular mass standard (size indicated).

Article Snippet: In a final chromatographic purification step, protein was loaded on a Bio-Gel HT hydroxylapatite column (Bio-Rad, Munich, Germany) equilibrated in buffer F. The eluate of a 12 mL gradient of 10–500 mm KPO4 in buffer F was collected in 0.3 mL fractions and assayed for PFK activity.

Techniques: Staining, Gradient Centrifugation