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

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.