Filtration of the cell lysate

A Milipore Pellicon XL 300 membrane actuated with a SciLog TANDEM 1081 peristaltic pump.

Centrifugation does not remove all of the cell debris, therefore a tangential flow ultrafiltration step with a 300 kDa membrane is performed. The retentate is collected. Instead of this device you can also use a simple dead-end sterile filter (0.22 µm poresize) but it is easier the way described above (the dead-end filter gets clogged after some time...).

With the cleared lysate the histidine affinity tag comes into play - want to know how?

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