This document describes how to calibrate a ZORBAX GF-250 column using Bio-Rad gel filtration standards.
This document describes a method to calibrate and test the proper operation of the Agilent ZORBAX GF-250 size-exclusion column. It utilizes molecular-weight markers from Bio-Rad gel-filtration standards, which allow calibration in the appropriate molecular-weight range for this specific column type.
The application highlights that these protein standards are separated by size with good linearity. The text notes that while the ZORBAX GF-250 achieves better separation of smaller molecules down to 1 Kdal, the GF-450 is designed for larger molecules up to 1 mdal.
| Column Part Number | 884973-901 |
|---|---|
| Column Dimensions | 250 mm length, 9.4.x ID |
| Mobile Phase Concentration | 200mM Sodium Phosphate |
| Flow Rate | 2 mL/min |
| Detection Wavelength | 254 nm |
| Temperature Condition | Ambient |
Bio-Rad gel-filtration standards are used for calibration.
The GF-450 achieves better separation of large molecules up to 1 mdal.