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Invitrogen™ ZOOM™ Urea
Description
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:
- 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.
- 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 (r) 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.
Specifications
Specifications
| Content And Storage | ZOOM™ IEF Fractionator • ZOOM™ Disks, one pack of each pH ZOOM™ Reagents • Urea • Thiourea • CHAPS • Carrier ampholytes pH 3-10 • Anode buffers • Bathode buffers Store all components at 4°C. |
| Shipping Condition | Room Temperature |
| Product Line | ZOOM |
| Form | Powder |
| Reagent Type | Urea |
| Quantity | 1 kg |
| Product Type | Urea |
Frequently Asked Questions (FAQs)
The ZOOM IEF Fractionator includes the following components:
Chamber Assembly Tube with Anode Reservoir
Spill Trough with Cathode Reservoir Lid
Sample Chambers (7)
Sample Chamber O-ring Seals, red (10)
Sample Chamber Caps with O-rings (7)
Cathode End Sealer
Anode End Sealer
Cathode End Screw Cap
Spacers, black (8)
Spares Box 1
Sample Chamber O-ring Seals (8)
Sample Chamber Caps with O-rings (7)
Spares Box 2
Cathode Chamber Seals (2)
Spacers, black (8)
We do not recommend heating protein samples containing urea over 37 degrees C as elevated temperatures cause urea to hydrolyze to isocyanate, which modifies proteins by carbamylation.
We recommend storing them at -80 degrees C. We do not recommend storing them at -20 degrees C
The recommended ampholyte concentration in the sample rehydration buffer is 0.5%.
*If you are loading 5-50 µg of protein (pure protein or crude lysate) per ZOOM strip, use 0.5% ampholytes in the sample rehydration buffer.
*If you are loading >50 µg of protein (crude lysate or fractionated sample) per ZOOM Strip, use 0.5-2% ampholytes in the sample rehydration buffer.
Note: Higher ampholyte concentration requires longer focusing times.
The maximum volume of the protein sample should at most be 1/6 of the final sample volume that will be added to the strip. A good general volume would be 5-10µL. 140 µL of sample diluted in Sample Rehydration buffer is used to rehydrate each ZOOM Strip for the standard rehydration time of one hour. For overnight rehydration, we recommend using 155 µL of diluted sample.
For Research Use Only. Not for use in diagnostic procedures.
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