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Two-dimensional (2D) gels separate proteins according to their isoelectric points (pI) and molecular weight. First dimension separation is accomplished using immobilized pH gradient (IPG) strips and high voltages to induce migration of proteins to their respective pI. IPG strips are thin plastic backed strips of dehydrated polyacrylamide. The polyacrylamide on the strip contains distinct regions of pH that can vary from narrow to extremely broad pH ranges. These strips are rehydrated with sample in a compatible buffer. After ample rehydration of the polyacrylamide a voltage gradient is applied to the strip. Voltage arcing through the sample forces the proteins in the acrylamide to migrate to the specific pH in which the net charge of the protein is zero- "focusing." The large array of IPG strips pH ranges allows us to narrow in on a pH range of interest. This facility can accommodate all pH ranges and strip sizes from 7cm to 24cm.
Our facility uses two isoelectric focusing instruments for the first dimension separation of proteins; Amersham/ GE Ettan IPGphor II and the Bio-Rad Protean IEF cell . This facility is capable of running all available sizes of IPG strips in large quantities.
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