GrainGenes Reference Report: JFC-47-3243
Reference
JFC-47-3243
Title
Prepartive fractionation of gliadins by electrophoresis at pH 3.1 (A-PAGE)
Journal
Journal of Agricultural and Food Chemistry
Year
1999
Volume
47
Pages
3243-3247
Author
Rumbo M Chirdo FG Giorgieri SA Fossati CA Anon CA
Abstract
Purification of gliadin subclasses has been difficult since they share many biochemical and physicochemical properties. In this report, the optimization of a preparative electrophoretic method to fractionate gliadins is described. Separation was performed in preparative 7% polyacrylamide gels at pH 3.1. The separation performance was tested using analytical electrophoresis at pH 3.1 and capillary electrophoresis. Preparative gels of different lengths were employed. Using 5-cm preparative gels, several fractions of alpha-, beta-, and gamma-gliadins and fast-mobility and slow-mobility omega-gliadins were collected in 40 h of separation. Resolution was maintained at a protein load of up to 30 mg in each run. The highest efficiency of recovery was achieved using aluminum lactate as the collecting buffer. Fractionation with 10 cm in length gels improved resolution but increased operation times. Gels of 2 cm in length did not separate alpha/beta- and gamma-gliadins efficiently but were useful to separate the two main fractions of omega-gliadins in shorter times. In conclusion, preparative electrophoresis at low pH allowed the separation of alpha-, beta-, gamma-, and omega-gliadin fractions from crude material under nondenaturing conditions
Keyword
aluminum
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GrainGenes Reference Report: JFC-47-3243
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GrainGenes is a product of the Agricultural Research Service of the US Department of Agriculture. | |||