The ReadyPrep sequential extraction kit is used to extract proteins of differing solubility from cell lysates. Based on the work of Molloy et al. (1998) and Herbert et al. (1998), the kit uses a series of three extraction buffers, each differing in their detergent and chaotrope concentrations (Rabilloud et al. 1997). The insoluble residues from each extraction step are further extracted with reagents of increasingly stronger solubilizing power.
The ReadyPrep sequential extraction kit:
- Yields fractions that are suitable for direct analysis by a number of methods, including 2-D electrophoresis
- Can be carried out in a single microcentrifuge tube, minimizing protein loss
- Results in manageable 2-D protein patterns for simplified protein identification and gel matching
- Generates fractions with high protein loads to enable visualization of low-abundance proteins
Molloy, MP, et al. (1998). Extraction of membrane proteins by differential solubilization for separation using two-dimensional gel electrophoresis. Electrophoresis 19, 837–844.
Herbert, BR, et al. (1998). Improved protein solubility in two-dimensional electrophoresis using tributyl phosphine as reducing agent. Electrophoresis 19, 845–851.
Rabilloud, T, et al. (1997). Improvement of the solubilization of proteins in two-dimensional electrophoresis with immobilized pH gradients. Electrophoresis 18, 307–316.