HOOK™ 6X His Protein Spin Purification kit allows for the rapid purification of soluble, 6X His tagged protein from bacterial cultures.
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 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.
Bacterial- PELB™ kit 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 (DNA & RNA) released during cell lysis. The Bacterial-PE LB™ lysis eliminates viscosity build-up, allowing effective clarification with lower centrifugal force. No sonication required.
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/50ml culture 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 ~250ml bacteria.
- Isolate >1mg His tagged proteins from ~50ml bacterial culture
- 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 bacteria