Standardization of protein position in silver-stained two-dimensional polyacrylamide gel electrophoresis

✍ Scribed by Dr. Dennis A. Johnston; Sylvia Capetillo; Louis S. Ramagli; Juan Guevara Jr; Douglas M. Gersten; Lewis V. Rodriguez

• Publisher: John Wiley and Sons
• Year: 1984
• Tongue: English
• Weight: 810 KB
• Volume: 5
• Category: Article
• ISSN: 0173-0835
• DOI: 10.1002/elps.1150050209

✦ Synopsis

Standardization of protein position in silver-stained two-dimensional poly acrylamide gel electrophoresis

Methods for standardization of silver-"stained" protein position in two-dimensional polyacrylamide gel electrophoresis are presented for their application to simultaneous multiple gel systems. Four methods are discussed: the use of a single gel per simultaneous run for standardization with either ( 1) known quantities of commercially prepared high-or low-molecular-weight standards or (2) a known biological preparation such as T4 phage coat proteins; the use of each gel per simultaneous run to standardize itself by (3) including ankternal carbamylation train such as creatine phosphokinase and using a one-dimensional gel pattern of proteins such as rat heart whole homogenate along the margin(s) or (4) identifying "marker" protein spots in test samples that occur in all gels in a given study and standardizing to relative position. Method four is discussed in detail and examples given of its use in a comparative study of proteins in spontaneous primary hepatocellular carcinomas occurring in mice, and its use compared to the other methods.