This application note describes the use of Agilent PLRP-S 100Å column for small scale high efficiency separation of synthetic protein fragment.
The document describes standard methods for small-scale peptide purification, specifically focusing on the separation of Acyl Carrier Protein (ACP) fragments. It highlights the use of PLRP-S 100A, a rigid polystyrene reversed phase media, to achieve high-efficiency separations for quantities in the micro-gram range.
The note details specific chromatographic conditions used for the separation, including column dimensions, eluent composition, gradient profile, flow rate, and detection wavelength. Results show the successful identification of ACP components and truncated sequences.
| Column Part Number | PL1512-3800 |
|---|---|
| Media Type | PLRP-S 100A |
| Particle Size | 8 um |
| Column Dimensions | 150 x 4.6 mm |
| Eluent A Composition | 0.1% TFA in 1 % ACN/99% water |
| Eluent B Composition | 0.1% TFA in 99 % ACN/1% water |
| Gradient Profile | 10-60% B in 20 min |
| Flow Rate | 1.0 mL/min |
| Detection Wavelength | UV, 220 nm |
To demonstrate the gradient purification of a synthetic acyl carrier protein fragment.
PLRP-S 100A, a rigid polystyrene (divinylbenzene) reversed phase media.
Eluent A is 0.1% TFA in 1 % ACN/99% water and Eluent B is 0.1% TFA in 99 % ACN/1% water.