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Polyacrylamide gel electrophoresis of small peptides

1984; Wiley; Volume: 5; Issue: 3 Linguagem: Inglês

10.1002/elps.1150050302

1522-2683

Michael H. P. West, Roy S. Wu, William M. Bonner,

Molecular Biology Techniques and Applications

Abstract Two polyacrylamide gel electrophoretic systems for the separation of small peptides are described. In both systems the samples are concentrated by a buffer discontinuity, and the peptides separated according to size and charge in 40–50 % polyacrylamide resolving gels. The acidic gel has an operative pH between 3.0 and 3.1 while the alkaline gel has an operative pH between 9.1 and 9.2. The lower limit of resolution is about 100 daltons for a peptide with one positive charge in the acidic system. The radioactive peptides are detected by autoradiography. Peptides with a free amino group can be fixed in these gels with formaldehyde, so that fluorographic procedures can also be used for increased sensitivity. These techniques enable precise comparison of non‐derivatized peptides from complete protein fingerprints and offer a simple one‐dimensional analytical method for protein comparison, as well as a general method for separating and analyzing small peptides.