The use of a fluorescence detector increases the sensitivity of the chromatographic analysis. In this Tech Note we evaluated the different sensitivity modes of the fluorescence detector with a fluorescein-labeled protein and compared it to the UV signal.
Fig. 1 Overlay of FITC-BSA fluorescence peak measured with different sensitivity modes (red line HIGH, green line MED, blue line LOW). 1.3 μg of sample 2 was injected.
Fig. 2A Overlay of FITC-BSA fluorescence (red) and UV 280 nm (blue) peak measured with HIGH sensitivity. 1.3 μg of sample 2 was injected.
Fig. 2B Overlay of FITC-BSA fluorescence (red) and UV 280 nm (blue) peak measured with MED sensitivity. 1.3 μg of sample 2 was injected.
Fig. 2C Overlay of FITC-BSA fluorescence (red) and UV 280 nm (blue) peak measured with LOW sensitivity. 1.3 μg of sample 2 was injected.
System configuration
| Method | FPLC |
|---|
| Mode | SEC |
|---|---|
| Substances | Fluorescein isothiocyanate isomer I (FITC), Bovine Serum Albumin (BSA) |
| Key words | SEC, FPLC, AZURA FPLC, fluorescence, BSA, FITC, fluorescein-labeled protein |
| CAS number | 3326-32-7, 9048-46-8 |
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