Rapid purification of EGFP, EYFP, and ECFP with high yield and purity
| Autor: | Shelley R McRae, Gillian Robin Bushell, Christopher L. Brown |
|---|---|
| Rok vydání: | 2005 |
| Předmět: |
Ammonium sulfate
Chromatography Lysis biology Elution Hydrophilic interaction chromatography Green Fluorescent Proteins biology.organism_classification Fluorescence Recombinant Proteins Green fluorescent protein Luminescent Proteins chemistry.chemical_compound Spectrometry Fluorescence Transformation Genetic Bacterial Proteins chemistry Escherichia coli Aequorea victoria Animals Triethylamine Biotechnology |
| Zdroj: | Protein Expression and Purification. 41:121-127 |
| ISSN: | 1046-5928 |
| Popis: | Most current high throughput purification procedures for the green fluorescent protein (GFP) suffer from poor yields and low purity. An improved purification procedure that delivers highly pure protein (>95% homogeneity) in high yields (>70% of the initial fluorescent protein content) has been developed. The purification procedure requires only two steps: the cell lysate is heated to 60 degrees C for 4 min in ammonium sulfate and triethylamine, followed by hydrophobic interaction chromatography using isopropanol during the elution phase. The resulting pure product exhibits the same fluorescence profile as the crude sample. This procedure has been demonstrated on three commercial variants of GFP from Aequorea victoria, enhanced green, enhanced yellow, and enhanced cyan fluorescent protein (Becton-Dickinson). The yield and purity of material are superior to other recently described methods. |
| Databáze: | OpenAIRE |
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