2D PAGE Analysis

2D electrophoresis is a powerful technique capable of simultaneously resolving 1000s of proteins in a single procedure.
For more than 20 years it was the core technology of proteomics until the recent rise of LC/MS/MS techniques.
It has some limitations in the analysis of extremely large proteins (>200 kDa), hydrophobic & membrane proteins and electrofocusing of proteins with pIs at the extremes (<3 or >11).

The typical analysis procedure involves the following steps:

  • Proteome preparation (critical step)
  • Isoelectric Focusing (on an Immobilized pH gradient - IPG)
  • 2nd Dimension SDS-PAGE
  • Gel staining (visible / fluorescent)
  • Image capture
  • Image analysis (Progenesis software)
  • Spot identification (MS-based analysis)

Depending on the stain used, the procedure can produce quantitative results (e.g. Coomassie, Sypro Ruby, DIGE).

Sample preparation is THE key step to the production of good quality, reproducible gels.

Pinnacle can provide guidelines and protocols for sample preparation depending on the source of your starting material (e.g. bacteria, fungi, plant or animal tissue).

If you would like to discuss sample preparation, are thinking of incorporating 2D analysis in a proposal or need advice about pricing feel free to contact the unit.