Q. What makes Edge 200 different than an ultracentrifuge?

A. Since the Edge 200 employs a centrifugal force for separation, it is often mistakenly compared to an ultracentrifuge. There are fundamental differences between the two instruments:

• Edge 200 performs the Centricollation™ process, which uses centrifugal force to accelerate a step-wise density extraction or “flotation” process.
  
  • Ultracentrifuges use centrifugal force to perform a gradient and banding process.
• Centricollation sequentially collects individual well-defined density supernatants or suspensions as its fractions.
  
  • An ultracentrifuge harvests pellets or non-defined bands as its fractions.
• The Centricollation process uses media of increasing density to sequentially extract or to float the specific particles.
  
  • Ultracentrifugation never changes density media.
• Edge 200 provides quality, uniform, NOT “fuzzy” separations by applying equal centrifugal force to the entire sample, using specially designed axial rotation.
  
  • Ultracentrifuges provide different forces to the sample particles (minimum force, maximum force, etc.) during a separation.
• Edge 200 preserves sample integrity better, since the sample stays at high centrifugal force for only 2-3 minutes for each fraction.
  
  • Ultracentrifuges subject samples to high centrifugal force for multiple hours.
• Edge 200 can be automated to perform and collect multiple extractions
  
  • An ultracentrifuge cannot.

Q. Can I use Gradispec reagents on my ultracentrifuge?

A. Yes you may, but we cannot guarantee and support the performance.
Our reagents are designed and optimized for use on the Edge 200 system. They are not designed for use on other systems.

Q. If I have an Edge 200, do I have to use Gradispec reagents or can I make my own?

A.

• Our reagents contain proprietary components other than sucrose.
• Our reagents are highly accurate in their concentration and composition.
• The quality and reproducibility of the reagents will directly affect the quality and reproducibility of the separation results.

Q. Why does Edge 200 only process one sample each time?

A.

• Edge 200 can provide 10 to 20 fractions within one to two hours. These well-defined fractions provide sufficient capacity for most down stream analyses.
• Edge 200 can provide well-defined separations of all subcellular compartments in a single, rapid process, while most other methods can only separate a specific compartment in a similar or greater timeframe.
• Once a fraction of interest has been identified, it takes only 10-12 minutes to isolate a similar fraction in subsequent experiments
• Edge 200 is focused on upstream sample separation and fractionation. At this stage, most research laboratories obtain limited samples (diseased or non-diseased, treated or control) from their sample sources.

Q. What makes Edge 200 different than other fractionation instruments?

A.

• Edge 200, using the Centricollation process is an embodiment of our force-enhanced suspension and defined-density extraction technology-Edge technology. It is currently the only instrument that can provide the process.
• Edge 200 system provides a non-denaturing density based separation, which is orthogonal to traditional separation methods, such as HPLC and electrophoresis, which separate samples based on their hydrophobicity, charge, molecular weight or PI.
• Edge 200 provides a more efficient way to identify low abundance proteins, functional proteins, and biomarkers by fractionating, enriching and concentrating subcellular compartments in a single process.
• Edge 200’s fractionation is focused on providing information about biological process and on the density of biological compartments in which biology occurs.
• Centricollation extraction process may start at, or jump to any density step of interest without the need for going through the intervening steps.
• Sample integrity of biological compartments is maintained, critical when determining early differences between normal and diseased samples.
• Groups of proteins or markers are identified within biologically relevant environments allowing the user to follow the biology throughout the disease process.
• Edge 200 is complementary, not competitive with almost all down stream analyses, such as, western blots, electrophoresis, mass spectrometry, and arrays.

Q. Does sucrose interfere with down stream analyses and how do I remove it?

A.

• Sucrose is a small molecule without charge. It is easily removed either by diluting the sample with next step analysis buffer and using a spin column with a 5kD cutoff, or by doing buffer exchange using 2DE buffer when 2DE is the next step.
• Sucrose does not interfere with SDS-Page or western blot analyses.
• Sucrose can be easily removed by ziptip for MALDI mass spec.
• For LC-MS/MS, sucrose will be eluted first at the beginning the gradient.