Filling of a Gel Filtration Column

The GPC column, which is intended for fractionation in laboratory scale, should have a ratio of bed diameter d to bed height H (Fig. 3.3B) of 1 50 to 1 200. 

Slurry the swollen, settled gel in about a half of its volume of buffer and degas under vacuum for 10 min. Connect the column outlet with tubing ending in a vessel at the level of the upper border of the column. Then mount the filling reservoir to the vertical column as shown in Fig. 3.3A. Fill the column up to one-fourth to one-third of its height with buffer and pour the gel slurry without bubbles in one portion into column and reservoir. Regulate the recommended hydrostatic pressure Ah (cf. Fig. 3.3 Table 3.2) by lowering the outlet. 

When the gel has settled, fill the column with buffer and carefully insert the upper adaptor. Move the adaptor down to the surface of the packed bed and push it further 1-3 mm into the GPC gel. Avoid air bubbles and let the outlets open to avoid compression of the gel. 

If columns without adaptors are used, stabilize the surface of the gel by a sheet of (glass fiber) filter paper. The surface of the bed should be without craters. 

The GPC columns with bubbles or clefts within the bed have to be emptied and packed again prior use. 

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