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A modification for the improved analysis of differential protein accumulation by two-dimensional polyacrylamide gel electrophoresis

✍ Scribed by Abul K. M. Ekramoddoullah; Yingchun Tan

Publisher: John Wiley and Sons
Year: 1998
Tongue: English
Weight: 93 KB
Volume: 9
Category: Article
ISSN: 0958-0344
DOI: 10.1002/(sici)1099-1565(199807/08)9:4<159::aid-pca399>3.0.co;2-f

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✦ Synopsis

Phenolic compounds present in conifer tissues interfere with the analysis of conifer proteins by twodimensional (2-D) gel electrophoresis, causing gel streaking and distortion of protein patterns. A protocol was established by modifying the Hochstrasser method and using the Millipore 2 D Investigator electrophoresis system. The modifications included substitution of N,N'-methylene-bis-acrylamide with piperazine diacrylamine as a cross-linking agent, dilution of the protein sample to a defined level of sodium dodecylsulphate, and a new way for the equilibration of the isoelectric focussing gel. The modified protocol greatly reduced gel streaking and background staining of conifer protein samples. The number of protein spots per gel was increased from 340 to 700. The well-resolved gel provided additional advantages for computer assisted image analysis.

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