Using 7 cm immobilized pH gradient strips to determine levels of clinically relevant proteins in wheat grain extracts
نویسندگان
چکیده
The aim of the work was to test a relatively simple proteomics approach based on phenol extraction and two-dimensional gel electrophoresis (2-DE) with 7 cm immobilized pH gradient strips for the determination of clinically relevant proteins in wheat grain. Using this approach, 157 2-DE spots were quantified in biological triplicate, out of which 55 were identified by matrix-assisted laser desorption/ionization - time of flight tandem mass spectrometry. Clinically relevant proteins associated with celiac disease, wheat dependent exercise induced anaphylaxis, baker's asthma, and food allergy, were detected in 24 2-DE spots. However, alcohol-soluble gliadins were not detected with this approach. The comparison with a recent quantitative study suggested that gel-based and gel-free proteomics approaches are complementary for the detection and quantification of clinically relevant proteins in wheat grain.
منابع مشابه
Isoelectric Focusing in Immobilized pH Gradient Strips Using the IPGphor Unit: Sample In-Gel Rehydration.
Isoelectric Focusing in Immobilized pH Gradient Strips Using the IPGphor Unit: Sample In-Gel Rehydration Angelika Görg, Oliver Drews, and Walter Weiss This protocol was adapted from "IEF in IPG Strips Using the IPGphor Unit," contributed by Angelika Görg, Oliver Drews, and Walter Weiss, Chapter 16, in Purifying Proteins for Proteomics (ed. Simpson). Cold Spring Harbor Laboratory Press, Cold Spr...
متن کاملEvaluation of Protocols Used in 2-D Electrophoresis for Proteome Analysis of Young Rice Caryopsis
In order to obtain a high-resolution electrophorogram of rice young panicle proteome, we evaluated various protocols commonly used in two-dimensional (2D) polyacrylamide gel electrophoresis (PAGE) of proteins, including gel staining protocol, pH range of immobilized pH gradient (IPG) strips and sample loading quantity. Results showed that a silver staining protocol using sensitized solution con...
متن کاملUse of zwitterionic type of detergent in isolation of Escherichia coli O56 outer membrane proteins improves their two-dimensional electrophoresis (2-DE).
Escherichia coli O56 were originally isolated from infected humans. Here it is reported that using the zwitterionic detergent (Zwittergent Z 3-14) to isolate outer membrane proteins (OMPs) from Escherichia coli O56 is suitable for their separation by two-dimensional electrophoresis (2-DE) using pH 3-10 immobilized pH gradient IPG strips (BIO-RAD).
متن کاملDryland Wheat Grain Yield and Yield Components as Affected by Slope Direction and Residue Rates
Limited information is available on the effect of residues rates and slope direction on dryland wheat (Triticum aestivum L.) yield. This study was carried out to determine the effects of residues rates and tillage direction on grain yield and yield components of the Sardary wheat in a dryland region in Zanjan. Five wheat residues rates (0, 25, 50, 75 and 100% surface cover) were applied and i...
متن کاملEffect of crop residue and nitrogen levels in yield and yield attributing traits of rice under Rice-Wheat Cropping System
A 3- years (2015, 2016 and 2017) field study was carried out at National Wheat Research Program, Bhirahawa, Rupandehi, Nepal to evaluate the influence of crop residues and nitrogen levels on rice. The experiment was conducted in split plot design with three replications. Main plots were two crop residue levels (with crop residues of 30 cm wheat stubble and without crop residues) and sub plots c...
متن کامل