蛋白質転写の実際と応用

抄録

Optimal performance in blotting applications depends on several factors which are divided into three functions. First, elution of proteins from the gel must be maximized. Second, eluted proteins should bind to the membrane completely and be accessible for further analysis. Finally, protein molecules must be retained on the membrane surface throughout analysis. Elution from the gel depends on the properties of each protein (e.g. solubility, molecular weight, isoelectric point). On the other hand, binding, retention and detection sensitivity depend on the membrane in large part. For optimization of protein blotting, a basic understanding of membrane structure and its implications for performance are very important.
In the second part, the “Rapid Immunodetection Method” eliminates the blocking step and reduces the washing steps dramatically. The sensitivity of detection was compatible to that observed with a conventional immunodetection method that included a blocking step and extended washing steps. Furthermore, the Rapid Immunodetection Method required less than 50% of the time required to complete the conventional immunodetection method.