Practical Protein Chemistry - A. Darbre 1989

Methods for Solid-Phase Amino Acid Sequence Analysis
Cleavage on a Solid-Phase Sequenator
Carbamoylation Reaction

At this stage, buffer no. 1 is most commonly used [27]. A non-aqueous DMF-based buffer (containing 2.5% triethylamine or N-methylmorpholine) has been recently proposed [43, 60]. In the DABITC–FITC analysis, good results are obtained using buffer no. 3 (Fig. 12.5). Non-aqueous or DMF-containing buffers are preferable to the previously suggested aqueous ones (since the non-aqueous medium provides a hydrophobic environment for the peptide attached to the polymer). These buffers are suitable for both polymeric and Glass Supports.

A 5% solution of FITC in acetonitrile is used; for simultaneous work with DABITC and FITC, both Reagents are dissolved in DMF (Fig. 12.5), as DABITC undergoes less Hydrolysis in this solvent.

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FIG. 12.5. An improved program for microscale solid-phase analysis using DABITC–FITC. Abbreviations: MeOH — methanol containing 0.2% propylamine; buffer — buffer no. 3 for the carbamoylation reaction; DABITC — 0.5% solution of DABITC in DMF; FITC — 5% solution of FITC in DMF; TFA — anhydrous TFA [52].



Last update: 06/08/2026

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