ศูนย์นวัตกรรมเทคโนโลยีหลังการเก็บเกี่ยว

A general method for two-dimensional protein electrophoresis of fruit samples

Diane Barraclough , David Obenland, William Laing and Tanya Carroll

Postharvest Biology and Technology Volume 32, Issue 2 , May 2004, Pages 175-181

2004

บทคัดย่อ

During experiments characterizing and identifying proteins from controlled atmosphere-stored apple and peachfruit,we optimized methods for the extraction and two-dimensional electrophoresis (2-DE) offruitproteins, using commercially available immobilized pH gradient strips for the first dimension. The method is relatively rapid with minimalhandlingof small amounts of sample, and has been reproduced successfully for 2-DE of a variety offruitand plant tissues in our labs. Critical factors forfruittissues include using acetone precipitation following incubation in a lysis buffer, and a long iso-electric focussing time. We have observed no interference to focussing from such troublesomefruitcomponents as soluble pectins, polyphenolics, or high-acidityfruit,using this protocol. In addition we have used the method with no modification, for a range offruittissues including low protein sources (apple and peach flesh), high lipid material (avocadofruitflesh) and high acidity lemon tissue. As the method in our hands is straightforward and robust, we recommend the method for routine 2-DE separations offruitsamples.