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

نویسندگان

  • Diane Barraclough
  • David Obenland
  • William Laing
  • Tanya Carroll
چکیده

During experiments characterizing and identifying proteins from controlled atmosphere-stored apple and peach fruit, we optimized methods for the extraction and two-dimensional electrophoresis (2-DE) of fruit proteins, using commercially available immobilized pH gradient strips for the first dimension. The method is relatively rapid with minimal handling of small amounts of sample, and has been reproduced successfully for 2-DE of a variety of fruit and plant tissues in our labs. Critical factors for fruit tissues 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 troublesome fruit components as soluble pectins, polyphenolics, or high-acidity fruit, using this protocol. In addition we have used the method with no modification, for a range of fruit tissues including low protein sources (apple and peach flesh), high lipid material (avocado fruit flesh) and high acidity lemon tissue. As the method in our hands is straightforward and robust, we recommend the method for routine 2-DE separations of fruit samples. Published by Elsevier B.V.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

A Simpler and Faster Version of Two-Dimensional Gel Electrophoresis Using Vertical, Mini SDS-PAGE Apparatus

We have modified one of the most useful methods of protein separation; namely, two dimensional gel electrophoresis (2-DE). This modified version of 2-DE is not only simpler and easier but also faster than all the currently available methods. In this method, isoelectric focusing is carried out in the first dimension using a vertical sodium dodecyl sulfate polyacrylamide gel electrop...

متن کامل

A Novel Spot-Enhancement Anisotropic Diffusion Method for the Improvement of Segmentation in Two-dimensional Gel Electrophoresis Images, Based on the Watershed Transform Algorithm

Introduction Two-dimensional gel electrophoresis (2DGE) is a powerful technique in proteomics for protein separation. In this technique, spot segmentation is an essential stage, which can be challenging due to problems such as overlapping spots, streaks, artifacts and noise. Watershed transform is one of the common methods for image segmentation. Nevertheless, in 2DGE image segmentation, the no...

متن کامل

O-17: Study of Sperm Protein Profile in Oigo- and Normozoospermic Patients with Varicocele Using Two Dimensional Eectrophoresis

Background: The aim of this study was to compare the sperm protein profile between oligo- and normozoospermic patients with varicocele. Materials and Methods: Sperm samples of 20 normozoospermic and 20 oligozoospermic patients with varicocele were characterized using two dimensional electrophoresis (group 1 and group 2, respectively). Differences in protein expression were established using gel...

متن کامل

Evaluation of Acetonitrile Deproteinisation of the Serum Samples for the Analysis of Drugs in Serum Using Capillary Zone Electrophoresis

Deproteinisation with acetonitrile (along with methanol or other reagents) is a useful and rapid technique in analysis of drugs or their metabolites in human serum. In this paper application of this simple technique in biopharmaceutical analysis using capillary electrophoresis (CE) is evaluated. Some drugs with different ionic and protein binding properties were selected and dissolved in human ...

متن کامل

بررسی بیان پروتئین‌های القاء شده برگ دو رقم گندم تحت تنش کادمیوم و جیوه با استفاده تکنیک الکتروفورز دو بعدی

Wheat is an important source of human food. Cadmium and mercury bind to sulfhydryl groups of structural proteins and enzymes and cause inhibition in activity and decrease in protein production or interfere with the regulation of the enzymes. To study the effect of protein expression under different levels of cadmium and mercury, the experiment was conducted in a completely randomized design wit...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:

دوره   شماره 

صفحات  -

تاریخ انتشار 2004