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2-Dimensional Electrophoresis

2-Dimensional Electrophoresis. 생명과학부 김 원. Proteomics Pathway. Proteomics Pathway. History of 2-DE. 1956 -paper & starch gel 1975 -Coupling of IEF & SDS-PAGE. IEF. +. SDS-PAGE. Sample preparation. Solubility, size, charge, isoelectric point

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2-Dimensional Electrophoresis

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  1. 2-Dimensional Electrophoresis 생명과학부 김 원

  2. Proteomics Pathway

  3. Proteomics Pathway

  4. History of 2-DE • 1956 -paper & starch gel • 1975 -Coupling of IEF & SDS-PAGE

  5. IEF + SDS-PAGE

  6. Sample preparation • Solubility, size, charge, isoelectric point • No universal method due to the diverse nature of samples. • Essential to minimize protein modifications. • Protease • Salt

  7. Tissue sample • Break up the tissue while it is still frozen or at liquid nitrogen. • Pestle, Mortar, Homogenizer • In cells, washed in phosphate-buffered saline (PBS) and solubilized in sample buffer • Ultracentrifugation-DNA, RNA 제거

  8. 2-Dimensional Electrophoresis 1D (IEF) Equilibration Identification 2D (SDS-PAGE)

  9. Troubleshooting

  10. Application of Proteomics

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