Staining protein in isoelectric focusing gels with fast green |
| |
Authors: | Ronald E Allen Kenneth C Masak Pamela K McAllister |
| |
Institution: | 1. Department of Food Science and Human Nutrition, Michigan State University, East Lansing, Michigan 48824 USA;2. Department of Animal Husbandry, Michigan State University, East Lansing, Michigan 48824 USA |
| |
Abstract: | A rapid, simple technique for staining proteins in isoelectric focusing polyacrylamide gels was demonstrated using fast green in 10% acetic acid. Fast green has the distinct advantage of not binding to ampholytes, thus staining only protein. Maximum staining was achieved within 5 min, and bands were visible after 3 to 6 h of destaining. Background stain removal, however, was not complete until 72 h after placing gels in a diffusion destainer. Gel quantitation was demonstrated with actin using fast green and Coomassie brilliant blue R-250. A standard curve prepared with fast green was linear from 0.5 to 8 μg of actin in contrast to Coomassie brilliant blue R-250 which provided linearity from 0.1 to 2.5 μg actin. Application of fast green staining to quantitation of α-actin from cultured muscle satellite cells has been demonstrated. |
| |
Keywords: | |
本文献已被 ScienceDirect 等数据库收录! |
|