The ZOOM™ IEF Fractionator offers a fast, reliable method to reduce sample complexity, enrich low abundance proteins, and increase the dynamic range of detection. Solution phase isoelectric focusing with the ZOOM™ IEF Fractionator provides reproducible separations in three hours. Fractionated samples are ready for further analysis by two dimensional gel electrophoresis (2DE), one dimensional gel electrophoresis (1DE), or two dimensional liquid chromatography/mass spectrometry (2D LC/MS).
The ZOOM™ IEF Fractionator System (Figure 1) consists of the following components:
• The ZOOM™ IEF Fractionator is built for safety, reliability, and durability. It includes easy-to-assemble sample chambers, in-built buffer chambers, buffer trough, removable electrode assembly, and safety lid.
• ZOOM™ Disks are pre-cast polyacrylamide gels that eliminate the need for manual preparation and minimize any chance of cross contamination. These immobilized buffered disks are pre-labeled, disposable, and designed for single use, ensuring consistent and reproducible fractionation. They resolve samples into as many as 7 fractions (using 8 disks of specific pH) to six fractions (using six disks of specific pH) from pH 3–12. Simply place each disk between the sample chambers in the ZOOM™ IEF Fractionator to allow separation in a specific pH range.
• ZOOM™ Reagents are proteomic grade reagents, including ZOOM™ 2D Protein Solubilizers urea, thiourea, CHAPS, and ampholytes, that ensure optimal conditions for sample preparation.
• The Basic Protein Kit, which includes ZOOM™ Discs and ZOOM Strips for IEF of basic proteins, focusing buffers, and solubilizer kits.
Sample Chamber and Fractionator Unit Set-up Procedure
Figures 4–6 illustrate assembly of the sample chambers and fractionator unit. The sample chamber and components fit together easily and offer a leak-proof seal. Simply place disks between the sample chambers in the ZOOM™ IEF Fractionator to allow separation in a specific pH range. For example, if you want to fractionate proteins between pH 4.6 and 5.4, flank a sample chamber with a ZOOM™ Disk of pH 4.6 and one of pH 5.4.
Downstream Processing by 2D Gel Electrophoresis
Following fractionation, the separated samples can be subjected to further analysis using narrow or broad range ZOOM™ Strips on the ZOOM™ IPGRunner™ System. Follow with second dimension analysis using neutral pH NuPAGE™ ZOOM™ Gels.