One-pot labeling and purification of peptides and proteins with fluorescein maleimide |
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Authors: | Eric Vivès Bernard Lebleu |
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Institution: | Institut de Génétique Moléculaire de Montpellier, CNRS-UMR 5124, BP5051, 1919 route de Mende, 34033 Montpellier cedex 1, France |
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Abstract: | Fluorescein labeling of peptides and proteins is required for numerous biophysical or biological experiments such as fluorescence microscopy, fluorescence resonance energy transfer (FRET) or fluorescence imaging. The commonly used strategy relied on the coupling of the dye reagent followed by a gel filtration to recover the labeled molecule. Here we report a simplified method for the labeling of peptides and proteins on a cysteine residue and their purification. The method is based on the precipitation of peptides and proteins in acetone, fluorescein maleimide being soluble in this solvent. The excess of dye is fully eliminated after a couple of acetone washes and the precipitated peptide or protein is readily recovered. |
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