Fig. 1

Direct measurement of hexokinase activity from normal and breast cancer serum after PEP protein separation and elution (60 min). After elution of proteins from the 2-D gel into the 384-well PEP plate, the eluted proteins were further transferred into a 384-well Master Plate with 50 μl refolding solution. 25 μl of the solution from the Master Plate was further transferred into a 384-well Enzyme Assay plate, 25 μl of hexokinase assay components were was added to each well, the increase of OD 340 nm by NADP reduction was measured in a spectrophotometer, the OD readings were taken at 0 and 60 min. the OD340 nm increase from 60 min. was calculated by subtracting the value from the 0 min. reading. Boxed fractions indicated samples of interest