HOOK™ 6X His Protein Purification kit allows for the 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 passing clarified lysate through prepacked columns.
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.
HOOK™ 6X His Protein Purification kit is 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.
HOOK™ 6X His Protein Purification kit is optimized to yield >10mg/250ml 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 small scale, rapid His protein isolation from bacteria we recommend our spin format kits.
- Isolate >10mg His tagged proteins from ~250ml bacteria
- No sonication required, supplied with lysis buffer and enzyme
- Nickel or Cobalt resins available
- For the isolation of 6X His tagged recombinant proteins from bacteria