Immunoblotting of gliadins separated by acid PAGE: Analysis of electrotransference conditions

RUMBO, MARTIN; CHIRDO, FERNANDO GABRIEL; AÑÓN, MARIA CRISTINA; FOSSATI, CARLOS ALBERTO

FOOD AND AGRICULTURAL IMMUNOLOGY

TAYLOR & FRANCIS LTD

Año: 1997 vol. 9 p. 135 - 139

0954-0105

Several transference buffers and conditions (time and voltage) were tested to establish a suitable procedure for immunoblotting of gliadins after acid polyacrylamide gel electrophoresis (A-PAGE). Ethanolic extracts of wheat flour were separated by A-PAGE, transferred to nitrocellulose membranes and detected using an anti-gliadin serum. The blot resolution was highly dependent on the buffer employed for electrotransference. The best results were obtained when a 10 mM-H3PO4/NaH2PO4 buffer, pH 2.7, was employed. All gliadin fractions were transferred using this buffer. Sharp bands were obtained and good resolution was achieved.