Biomass either provided by you or alternatively prepared by us if using service D.
1. Preparation of the „Inclusion Bodies“ (IB)
- Disintegration of bacteria by lyses buffer and sonification .
- Accumulation of „IBs“ by several centrifugation steps .
- Check of „IB- preparation“ by SDS/PAGE and Comassie stain.
2. Solubilisation of „Inclusion Bodies“
- Solubilisation of „IBs“ .
- Check of „IBs“ by SDS/PAGE and Comassie stain
Note: Further measurements concering the purification depend on the protein of interest and the information available.
The following features influence the subsequent proceeding.
- existence of an established purification protocol,
- the protein is fused to a “tag”,
- the native protein is a dimer, or
- there has to be established a new protocol.
In case of 3. the next step is the dimerization reaction followed by a gel filtration to separate monomers from dimers.
In case of 4. the process is continued with ion exchange chromatography (e.g. anion/cation).
- The total yield of purified protein.
- A detailed report sheet.
Dependent on the protocol used.