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EMF Protein Analysis Flow Chart

Isolated Protein. Gel protein spot (1D / 2D). Blotted on PVDF membrane. Figure I. EMF Protein Analysis Flow Chart. Biological material (Tissue, Cells, Body fluids). HPLC/ Purification. Gel electrophoresis. Digestion/ cleavage. Digestion/ cleavage. Digestion/ cleavage.

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EMF Protein Analysis Flow Chart

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  1. Isolated Protein Gel protein spot (1D / 2D) Blotted on PVDF membrane Figure I EMF Protein Analysis Flow Chart Biological material (Tissue, Cells, Body fluids) HPLC/ Purification Gel electrophoresis Digestion/ cleavage Digestion/ cleavage Digestion/ cleavage Edman degradation (sequence) 500 fmol Amino Acid Analysis 1 - 10 pmol Peptide mixture Edman degradation (sequence)  500 fmol Desalting (ZipTip) or Nanodesalting Proposed Addition HPLC fractionation Fractionated peptide MALDI-TOF (mass map)  50 fmol LC-ESI-MS (mass map) 1 - 5 pmol Nano ESI-MS (mass map) 0.5 - 1 pmol LC-ESI-MS/MS (MS scans plus MS/MS scans) mass map + sequence tag Q-Tof Ultima 50 - 500 fmol PSD MALDI-TOF (sequence tag)  100 fmol Nano ESI-MS/MS (sequence tag) 0.5 -1 pmol Edman degradation (sequence)  300 fmol PSD MALDI-TOF (sequence tag)  100 fmol Nano ESI-MS/MS (sequence tag) 0.5 - 1 pmol Database searches (SEQUEST, Mascot, PROWL, Protein Prospector, etc.)

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