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United States Department of Agriculture

Agricultural Research Service

Proteomics
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1 - Preparation of Proteins for Mass Spectrometry
2 - Mass Spectrometric Analysis
3 - High Performance Liquid Chromotography (HPLC)
4 - Molecular Modeling
Preparation of Proteins for Mass Spectrometry

Wheat Quality Research

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Endosperm
Step 1. Wheat grains are harvested from plants grown in the greenhouse and endosperm (arrow) is collected.
2D Gel
Step 2. Proteins are extracted and separated by two-dimensional (2D) gel electrophoresis and stained with Coomassie blue.

 

Computer
Step 3. The 2D gels are scanned and the images are matched and analyzed using sophisticated computer software.
Spot Density Graph
Step 4. The computer software quantifies gel spots and makes it possible to compare protein profiles during grain development.

 

Spot Picker
Step 5. The computer generated spot coordinates are used to program a robotic spot picker that punches the spots from the 2D gels and places them into 96 well plates.
Digest Pro
Step 6. The spots in the 96 well plates are processed in a digestion robot to yield peptides for analysis by mass spectrometry.

Reference

Hurkman, W.J. and Tanaka, C.K. 2004. Improved methods for separation of wheat endosperm proteins and analysis by two-dimensional gel electrophoresis. J. Cereal Sci. 40: 295-299.

 

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Last Modified: 5/1/2006
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