Capacities of human serum and plasma 225

Lynn A. Echan, Hsin-Yao Tang, Nadeem Ali-Khan, KiBeom Lee and David W. Speicher

5.1 Introduction 226

5.2 Materials and methods 227

5.2.1 Serum/plasma collection 227

5.2.2 MARS 228

5.2.3 Multiple affinity removal spin cartridge 228

5.2.4 Microscale solution IEF (MicroSol IEF) (ZOOMâ„¢-IEF) fractionation 228

5.3 Results 220

5.3.1 Depletion of major proteins to enhance detection of lower abundance proteins 220

5.3.2 Evaluation ofhigh-abundance protein removal using 2-DE 222

5.3.3 Specificity of major protein depletion 223

5.3.4 Impact of Top-6 protein depletion on detection of lower abundance proteins using 2-D gels 225

5.3.5 Combining Top-6 protein depletion with microSol IEF prefractionation and narrow pH range gels 225

5.3.6 Analysis of Top-6 depleted serum and plasma using protein array pixelation 228

5.4 Discussion 230

5.5 References 234

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