centrifugation at 105,000 g for 1h endonuclease Cultured animal cell line Tissue, E. Coli, yeast.. washing with low salt buffer rounded mechanically under liquid nitrogen assing through a syringe needle onication ounce homogenization urea (7 M), thiourea (2M), CHAPS (4% w/v) DTT (15 mM) protease inhibitor cocktail Tris (40 mM) GSH (1 mM) EDTA (0.5 Mm) protease inhibitor cocktail or Serum Diluted with separation media final protein concentration of 0.5~6.0 mg FFE 1500V, 25 mA, ~33 min max. 70 fractions (pH 2.5~11) 1D PAGE
washing with low salt buffer. urea (7 M), thiourea (2M), CHAPS (4% w/v) DTT (15 mM) protease inhibitor cocktail. or. Tris (40 mM) GSH (1 mM) EDTA (0.5 Mm) protease inhibitor cocktail. grounded mechanically under liquid nitrogen passing through a syringe needle sonication - PowerPoint PPT Presentation
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centrifugation at 105,000 g for 1hendonuclease
Cultured animal cell lineTissue, E. Coli, yeast..
washing with low salt buffer
grounded mechanically under liquid nitrogenpassing through a syringe needle sonication Dounce homogenization