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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

2-Dimensional Electrophoresis

생명과학부

김 원




History of 2 de
History of 2-DE

  • 1956

    -paper & starch gel

  • 1975

    -Coupling of IEF & SDS-PAGE


IEF

+

SDS-PAGE


Sample preparation
Sample preparation

  • Solubility, size, charge, isoelectric point

  • No universal method due to the diverse nature of samples.

  • Essential to minimize protein modifications.

  • Protease

  • Salt


Tissue sample
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


2 dimensional electrophoresis1
2-Dimensional Electrophoresis

1D (IEF)

Equilibration

Identification

2D (SDS-PAGE)




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