Affinity chromatography by His-Tag is one of the most widespread purification techniques for recombinant proteins. In many cases it requires an additional cleaning/polishing step. This application highlights the possibility of combining two subsequent chromatography protocols without manual interaction using the AZURA Bio LC system.
Chromatogram of the two-step 6xHis-GFP purification; 280 nm UV signal
Chromatogram of the two-step 6xHis-GFP, GFP detection with 395 nm UV signal
SDS-PAGE of two-step 6xHis-GFP purification
| Method | FPLC |
|---|
| Mode | SEC |
|---|---|
| Substances | Green fluorescence protein (GFP) with 6xHis - Tag |
| Key words | GFP, His-Tag, IMAC, FPLC, affinity, SEC, buffer exchange |
| CAS number | - |
AZURA® Detector MWD 2.1L Multiwavelength detector MWD 2.1L, without flow cell 190 - 700 nm, incl. test cell
Article No.: ADB01
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