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Figure 1 | Proteome Science

Figure 1

From: Evaluation of three high abundance protein depletion kits for umbilical cord serum proteomics

Figure 1

Comparison of depletion efficiency and repeatability of different kits on umbilical cord serum proteins samples by 1D-SDS-PAGE. Ten micrograms of crude, depleted or bound protein were loaded onto each lane and separated by 4-12% polyacrylamide gels. Proteins were visualized by Coomassie Blue staining. For examination of repeatability in umbilical cord serum depletion, the immunoaffinity columns (5185-Agilent and PROTIA-Sigma) were run three consecutive times with re-equilibration between depletion processes. However, for Blue Albumin and IgG Depletion Kit (PROTBA, Sigma-Aldrich, Saint Louis, MO, USA), serum samples were tested in two independent columns since its depletion efficiency was low even with a new column. A) Blue Albumin and IgG Depletion Kit (PROTBA, Sigma-Aldrich, Saint Louis, MO, USA) in umbilical cord serum depletion. B) Blue Albumin and IgG Depletion Kit (PROTBA, Sigma-Aldrich, Saint Louis, MO, USA) in adult venous serum depletion. C) Multiple Affinity Removal System (5185, Agilent, Santa Clara, CA, USA) in umbilical cord serum depletion. D) Immunoaffinity Albumin and IgG Depletion Kit (PROTIA, Sigma-Aldrich, Saint Louis, MO, USA) in umbilical cord serum depletion. Lanes: M: Prestained protein marker (Fermentas; 250, 130; 95, 72, 55, 36, 28, 17); C: Crude serum; D: Depleted serum; B: Bound proteins; D1-3: Depleted umbilical cord serum proteins of the first, second and third depletion process. B1-3: Bound proteins eluted from the column after the first, second and third depletion process. Dp: Depleted Proteins pooled by the three depletion processes. Bp: Bound proteins pooled by the three depletion processes Closed arrows: Protein band indicating albumin; Open arrows: Protein band indicating IgG; * heavy chain, and ** light chain.

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