This document introduces the key technology in protein analysis-size exclusion chromatography (SEC), and illustrates the principle, characteristics and application of SEC with Agilent ProSEC 300S column as an example
This document details the use of size exclusion chromatography (SEC) for protein analysis, highlighting the importance of minimizing protein interactions with column packing material. It presents an experiment using a series of globular proteins to validate the performance of the ProSEC 300S column.
| Column Dimensions | 300 x 7.5 mm |
|---|---|
| Eluent Composition | 0.3M NaCl, 50mM KH2PO4-K2HPO4, pH 6.8 |
| Flow Rate | 1.0 mL/min |
| Injection Volume | 20 pL |
| Temperature | 25 °C |
| Detection Wavelength | 280 nm |
The calibration used Thyroglobulin, y-Globulin, Bovine serum albumin, Ovalbumin, Carbonic anhydrase, Myoglobulin, Cytochrome C, and Bacitracin.
The calibration curve shows a linear relationship between the log of the molecular weight and the retention time.