G-Biosciences' HOOK™ 6X His Protein Purification (Bacteria) is a complete kit for the purification of soluble, 6X His tagged protein from bacterial cultures
G-Biosciences' HOOK™ 6X His Protein Purification (Yeast) is a complete kit for the 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 IMAC by passing the clarified lysate through prepacked columns. Yeast PE LB™ is based on organic buffering agents that utilize a mild non-ionic detergent and a combination of 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.
G-Biosciences' HOOK™ 6X His Protein Purification kits (bacteria or yeast) are available with either nickel chelating immobilized metal affinity chromatography (IMAC) columns or cobalt chelating IMAC columns. 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. The HOOK™ 6X His Protein Purification kit are optimized to yield >10 mg of soluble His tagged protein, with a purity of 80-90%, dependent on expression levels, resin type, conformation and solubility characteristics of the protein. Suitable for 5 isolations. For small scale, rapid isolation, use G-Biosciences' HOOK™ 6X His Protein Spin Purification kits (bacteria or yeast), which are optimized to yield about 1 mg of soluble His tagged protein, with a purity of 80-90%.
The bacteria are first lysed with Bacterial PE LB™ and PE LB™-Lysozyme to release total soluble protein, whilst maintaining the structure and activity of the protein. The 6X His tagged protein is purified by IMAC by passing clarified lysate through prepacked columns. Bacterial- PELB™ has been developed for the extraction of soluble proteins from bacterial cells. It is a proprietary improvement on the lysozyme based lysis, which allows extraction of soluble proteins and concurrent removal of nucleic acids released during cell lysis. The Bacterial-PE LB™ lysis eliminates viscosity build-up, allowing effective clarification with lower centrifugal force.