HOOK™ 6X His Protein Spin Purification kit allows for the rapid purification of soluble, 6X His 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 6X His tagged protein is purified by immobilized metal affinity chromatography (IMAC) by adding 0.5ml immobilized metal affinity resin to the clarified lysate. The resin is transferred to a convenient spin column, where it is rapidly washed and the 6X His protein is eluted with an imidazole buffer.
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™ 6X His Protein Spin Purification kit is available with either nickel chelating resin or cobalt chelating resin for the immobilized metal affinity chromatography. Cobalt chelating resin has a lower binding affinity for 6X His tags, compared to nickel chelating resin, which results in less non-specific binding and may result in slightly lower yields.
HOOK™ 6X His Protein Spin Purification kit is optimized to yield ~1mg of soluble His tagged protein, with a purity of 80-90%, dependent on expression levels, resin type, conformation and solubility characteristics of the protein.
For larger scale His protein isolation from bacteria we recommend our gravity flow kits for >10mg protein from yeast.
- Isolate >1mg His tagged proteins from yeast
- No sonication required, supplied with lysis buffer and enzyme
- Nickel or Cobalt resins available
- Spin column format
- Spin column format for the isolation of 6X His-tagged recombinant proteins from yeast