User:Maria Briscione/Notebook/Chem 571/2012/02/08

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Objective

Fluorescently label cysteine residues on BSA

Description

Procedure

  • 406 microliters of the phosphate buffer (pH 7.04)
  • 2.9 mg of 5-iodoacetamido flourescin
  • 2.776 mL BSA from Stock
  • BSA concentration - 13.086 ┬ÁM
  • let reaction sit for at least two hours in dialysis bag
  • dialysis over a week, water was changed daily

Data

Notes

  • Ideal ph is between 7 and 8
  • deprotonates at this pH
  • labeling was completed in order to determine binding of the cysteine to the dye
  • this procedure also allows us to determine concentration


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