•High throughput protein expression protocols •

Expression clones are characterised by high throughput protein expression experiments. Bacteria cultivation and protein expression is generally done in microtitire plates. We analyse whole cellular protein (including inclusion bodies) and soluble cellular protein, and we perform small scale protein purification experiments under denaturating and non-denaturating conditions. Protein purification is performed in 96-well microtitre plate format, either manually or on a automated pipetting robot. We use Ni-NTA or glutathtion affinity purification for His-tag or GST-tag fusion proteins.

The first protocol describes the protein expression in microtitre plates:

1. Protein expression in microplates

According to the different options, a set of protocols exists to proceed from protocol 1:

2. Soluble and whole cellular protein extracts.
3. Ni-NTA affinity purification (non-denaturing, manually)
4. Glutathion affinity purification (non-denaturing, manually)
5. Ni-NTA affinity purification (denaturing, manually)
6. Non-denaturing Ni-NTA affinity purification (robot)
7. Non-denaturing glutathion affinity purification (robot)

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© 2003 by V. Sievert, Konrad Büssow last changed 12 Sep 2006