Gang-hua Lang, Mika Ohba, Shinichi Kawamoto, Koichi Yoza, Tatsuya Moriyama and Kazumi Kitta
Food Sci. Technol. Res. 2010. Vol 16.

Overview

  • Keywords: Rice (Oryza sativa), RAG2, α-amylase/trypsin inhibitor, recombinant protein, polyclonal antisera, processed food.
  • Aim of the study: Allergen identification in processed food
  • Application: SDS-PAGE analysis
  • Sample type: Cereals
  • Material: FastPrep-24™ instrument, 2 ml Lysing Matrix D tubes, Lab Mixer
  • Buffer: Buffer A (30 mM Tris-HCl, pH 8.0 supplemented with 1 M NaCl)

Protocols & Parameters

  1. Grains of various cereals were crushed into flour with a Lab Mixer
  2. 100 mg of grain flour was transferred to a 2 ml Lysing Matrix D tubes
  3. 1 ml of Buffer A was added to the tubes
  4. After brief vortexing, the tubes were incubated on ice for 1 h
  5. The tubes were then set up in a FastPrep® System at speed 6.0 for 40 sec
  6. After centrifugation at 15,000 x g, 4°C for 10 min, the supernatant was dispensed into several tubes and stored at −30°C

Conclusion

  • The use of FastPrep® system in combination with lysing matrix D allowed isolation and characterization of the main allergens in rice grains.