Calibration standards

Table 8.3.8 provides a compilation of commercially available molecular size calibration standards and their suppliers. Some of these are available as kits (e.g., from Amersham Pharmacia Biotech). Protein standards should be prepared in the GF buffer that will be used in the molecular size determination. To prolong the life of the column, samples should be filtered through a protein-compatible filter prior to injection. The exact concentration to be prepared depends on the additives present (e.g., sucrose as stabilizer), detector sensitivity, and the zone broadening of the column. Concentrations of proteins in the range from 1 to 5 mg/ml are frequently used for calibration. It is possible to mix several standards to calibrate the separation range of interest in a few runs (it is seldom necessary to calibrate the entire separation range of the column).

Table 8.3.8 also lists some dextran fractions that are useful for calibrating the column in hydrodynamic volume, which makes comparing solutes of different shape easier.

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