Influence of gel dimensions on resolution and sample throughput on two-dimensional gels
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To achieve high throughput and economical format of 2-D PAGE, comparison between gel size and resolution was conducted on human breast carcinoma cell line (MCF-7/AZ) proteins. SDS gel length showed a weaker influence of separation length on resolution in the second dimension, and there was little benefit of separation distances greater than 15 to 19 cm. IPG strip separation distances were very important with dramatic increase in resolution of longer gels compared with smaller gels, and maximal resolution was obtained using 18-and 24-cm IPG strips. Loading optimal amount of proteins on 2-D gels can also increase the number of detected spots. Therefore, taken together, compromise 2-D gels are crucial for higher capacity and higher throughput.
Key wordstwo-dimensional polyacrylamide gel electrophoresis immobilized pH gradient (IPG) strip length SDS gel length/high throughput resolution
- 2-D PAGE
two-dimensional polyacrylamide gel electrophoresis
immobilized pH gradient
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