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. 2016 Jun 15;6(8):816–826. doi: 10.1002/2211-5463.12091

Figure 2.

Figure 2

Preparation of secreted proteins. (A) A scheme of the experimental design comprising sample preparation and iTRAQ labeling. CM was collected and fractionated with a series of cut‐off spin columns for the removal of transferrin (76 kDa) derived from the ITS ™ supplement. ‘Low’ and ‘high’ fractions were fractionated using a series of spin columns. Each fraction of CM was labeled with a different iTRAQ ® reporter as indicated. (B) Evaluation of secreted proteins by SDS/PAGE. Equivalent amounts of secreted proteins from each sample (D2, D4, and D8) were subjected to SDS/PAGE and subsequently stained with SYPRO ® Ruby. The arrowhead indicates bands corresponding to transferrin. D2, day 2; D4, day 4; D8, day 8; M, molecular weight marker.