Problems encountered in measuring erythrocyte pyrimidine 5'-nucleotidase activity |
| |
Authors: | J D Torrance M S Wulfsohn W Mills |
| |
Institution: | Research Institute of the Palo Alto Medical Foundation, 860 Bryant Street, Palo Alto, California 94301 USA |
| |
Abstract: | An improved method of two-dimensional electrophoresis that allows unequilibrated first-dimension gels to be loaded and electrophoresed on second-dimension gels twice the length used in the O'Farrell technique has been developed. Normally, the electrophoresis of unequilibrated first-dimension gels on long second-dimension gels with the resolving gel set at pH 8.8 results in poor resolution of low-molecular-weight proteins. Adjusting the pH of the resolving gel to pH 8.3 maintains the low-molecular-weight proteins in a stacked configuration during their migration through the length of the 10% acrylamide gel. Utilization of a 10 to 20% exponential polyacrylamide gradient in a resolving gel set at pH 8.3 separates these low-molecular-weight proteins with excellent resolution. Electrophoretic resolution of protein spots in resolving gels set at pH 8.8 is not as sharp as in gels set at pH 8.3, and resolution progressively deteriorates in gels set at higher pH values. |
| |
Keywords: | To whom correspondence should be addressed at Department of Nutrition and Food Science 56-225 Massachusetts Institute of Technology Cambridge Mass 02139 |
本文献已被 ScienceDirect PubMed 等数据库收录! |