Practical Protein Chemistry - A. Darbre 1989

Methods of solid-phase amino acid sequencing
Cleavage on a solid-phase sequencer
Column

The design of the instrument has been described previously [27—29, 35].

Both ends of a thin Glass tube (length 230 mm, internal diameter 2.5 mm) are polished to obtain a flat end surface. The connections are made of Teflon and Kel-F [27]. The column is placed in a heated aluminum block thermostated at 45 °C.

12.5.1.1. Microcolumn. For the analysis of Peptides in amounts <20 nmol, downsized columns (e.g., 15X2 mm) packed with peptidyl-glass beads mixed with glass filler are used. A column design made of Teflon tubing (Fig. 12.4), whose internal volume corresponds to The amount of peptidyl-glass used, is known. A porous Teflon filter (pore size 2 µm) is placed under the tapered lower end of the tube, resting in an adapter that seals the end of the column. The dead volume at the top of the column is minimized by inserting a short piece of narrow Teflon tubing. The assembled column is placed in a glass thermostat (Fig. 12.6).

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FIG. 12.4. Column for peptide analysis at the micro-level. The column material is Teflon tubing, the internal volume of which depends on the amount of support used. A narrow Teflon tube is inserted into the upper part of the column, and the lower end of the column is closed with a porous Teflon filter with a pore diameter of 2 µm (Zitex, Chemplast Inc.). The column is placed in a thermostated glass tube (see Fig. 12.1) [52, 69].