HOOK™ GST Protein Spin Purification kit allows for the rapid purification of soluble, GST tagged protein from yeast cultures. The yeast are first lysed with Yeast PE LB™ and LongLife™ Zymolyase® to release total soluble protein, whilst maintaining the structure and activity of the protein. The GST tagged protein is purified by affinity chromatography by adding 0.5ml Glutathione resin to the clarified lysate. The resin is transferred to a convenient spin column, where it is rapidly washed and the GST protein is eluted.
Yeast-PE LB™ is useful for extraction of soluble proteins from yeast cells. Yeast PE LB™ is a proprietary improvement on the Zymolyase® based spheroplast preparation and extraction of soluble proteins from yeast cells. Yeast PE LB™ is based on organic buffering agents that utilize a mild non-ionic detergent and a proprietary combination of various salts and agents to enhance extraction and stability of proteins. A ready-to-use Zymolyase® preparation is also provided. Depending on application, additional agents such as protease inhibitors may be added into Yeast PE LB™. The proprietary combination of this reagent provides a simple and versatile method of yeast protein extraction. Yeast PE LB™ eliminates the need for laborious glass bead lysis of yeast cells.
HOOK™ GST Protein Spin Purification kit is optimized to yield ~1mg of soluble GST tagged protein, with a purity of 80-90%, dependent on expression levels, resin type, conformation and solubility characteristics of the protein.
For larger scale GST protein isolation from yeast we recommend our gravity flow kits for >10mg protein from yeast.
- Isolate >1mg GST tagged proteins from ~50ml yeast culture
- No sonication required, supplied with lysis buffer and enzyme
- Spin column format
- Spin column format for the isolation of GST-tagged recombinant proteins from yeast