Abstract
A novel technique to selectively analyze prelabeled peptides by isoelectric focusing (IEF) is presented. The conditions are described for biotinylation of peptides, their separation in polyacrylamide gels by IEF, and their fixation to the gel matrix with glutaraldehyde. The gels are developed by a color reaction catalyzed by an avidin-coupled enzyme. The technique is suitable for peptides with at least one free amino group of guanidino group after N-terminal biotinylation. The presence of other peptides or proteins does not interfere with the detection. The sensitivity is below 10 pmol, representing a 1000-fold improvement over existing techniques for analyzing low molecular weight peptides by IEF.
Original language | English |
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Pages (from-to) | 96-103 |
Number of pages | 8 |
Journal | Analytical Biochemistry |
Volume | 171 |
Issue number | 1 |
DOIs | |
Publication status | Published - 15 May 1988 |
Externally published | Yes |
Keywords
- angiotensin II
- avidin-enzyme staining
- biotinylation
- isoelectric focusing
- peptide
- polyacrylamide gel