Selected article for: "mm tcep and room temperature"
Author: Wilton T. Snead; Wade F. Zeno; Grace Kago; Ryan W. Perkins; J Blair Richter; Chi Zhao; Eileen M. Lafer; Jeanne C. Stachowiak
Title: BAR scaffolds drive membrane fission by crowding disordered domains
  • Document date: 2018_3_4
    • ID: drqseaaa_51
    Snippet: Proteins were labeled using amine-reactive, NHS ester-functionalized dyes (Atto-Tec) in 25 mM HEPES pH 7.35, 150 mM NaCl, 5 mM TCEP buffer. The concentration of dye was adjusted experimentally to obtain the desired labeling ratio of 0.5-1 dye molecules per protein, typically 2-5 times molar excess of dye. Reactions were performed for 20-30 min at room temperature and labeled protein was separated from unconjugated dye using Princeton CentriSpin-2.....
    Document: Proteins were labeled using amine-reactive, NHS ester-functionalized dyes (Atto-Tec) in 25 mM HEPES pH 7.35, 150 mM NaCl, 5 mM TCEP buffer. The concentration of dye was adjusted experimentally to obtain the desired labeling ratio of 0.5-1 dye molecules per protein, typically 2-5 times molar excess of dye. Reactions were performed for 20-30 min at room temperature and labeled protein was separated from unconjugated dye using Princeton CentriSpin-20 size exclusion spin columns (Princeton Separations).

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