Highly resolving two-dimensional gels for protein sequencing.

摘要: Abstract Two-dimensional (2D) PAGE, using carrier ampholytes for the first-dimension separation, has provided a tool simultaneous analysis of cellular proteins. To extend utility 2D PAGE to preparative level, we have investigated use immobilized pH gradients (IPG) separation. The results obtained indicate that as much 1 mg protein can be loaded onto single IPG gel without loss resolution. Mutant polypeptides previously detected in ampholyte-based gels were equally detectable IPG-based gels. With several hundred isolated, from few 10 gels, sufficient amount sequencing with current technology. We therefore conclude greatly enhances prospects large-scale proteins development gel-related data bases and identification new polypeptide gene products, attendant implications genome effort.