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GELFrEE® 8100 Fractionation Station – Request a demo

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Protein Fractionation System*

Why choose GELFrEE?

In proteomics mass spectrometry analysis, protein mixtures are so complex that the low abundant proteins, the main ones of interest, are lost among the highly abundant signals. Excising protein bands from preparative slab gels can result in little or no protein. Hence, effective fractionation of complex samples for in-depth proteomic analysis by mass spectrometry is critical.

Expedeon GELFrEE® enables molecular weight-based fractionation of intact proteins which are recovered in liquid-phase fractions. It gives high protein recovery rate thus allows in-depth study of the whole proteome.

How it works?

GELFrEE®  is “Gel-eluted Liquid Fraction Entrapment Electrophoresis”. Each of the eight GELFrEE® 8100 Cartridge channels consists of a precision-cast gel column surrounded by pipette-accessible sample loading and fraction collection chambers. Samples are mixed with the pre-formulated Sample Buffer and pipetted into the sample loading chamber.

When electric current is applied, charged molecules in the channel migrate first, focus in the stacking gel, and then separate according to their electrophoretic mobility or, when using SDS, molecular weight. Smaller proteins move more quickly and elute into the fractionation collection chamber, where they are trapped in a defined liquid volume. At predefined intervals, the instrument will then automatically pause for easy removal of the liquid fraction with a pipette. For collection of the next size-based fraction, the sequence is restarted and the process continued.

1D gel of S. cerevisiae fractions prepared using the GELFrEE® 8100 System.

A 200µg aliquot of yeast was fractionated into 12 fractions using the GELFrEE® 8100 system. 1D gel analysis with silver staining was used to visualize the results.

Basepeak chromatogram of digested fraction from GELFrEE® 8100 System.

A yeast sample, S. cerevisiae, was fractionated into 12 fractions. The resulting fractions were acetone precipitated to remove SDS, digested using trypsin, and analyzed using nanoLC-MS/MS.

Features

The compact, bench-top GELFrEE® 8100 Fractionation Station provides specified voltage to each of eight independent channels and pauses for fraction collection according to user programming. Using the touch screen interface, the fractionation sequence is defined or selected, permitting broad mass range analyte fractionation or target protein enrichment in accordance with the users requirements. During operation, the touch screen interface provides feedback on the status and current/voltage that is applied for each channel.

  • Intact protein molecular weight fractionation, isolation, and purification
  • Allows in-depth study of whole proteome
  • Liquid-phase recovery without band or spot cutting
  • Broad mass range fractionation up to 500 kDa
  • Up to eight samples processed in parallel
  • Programmable fractionation for isolating and purifying targeted proteins
  • High protein recovery (>80%)
  • High reproducibility (>15% CV)
  • Sampling unbiased by hydrophobicity,  isoelectric point (pI)
  • High loading capacity (>5X more than a 1D gel)
  • Proteins are recovered intact, for complete characterization

 

Application

  • Simplify and reduce the dynamic range of complex protein mixtures for bottom-up discovery proteomics using LC-MS/MSApplications
  • Fractionate and recover proteins intact for top-down proteomics
  • Isolate and enrich user-selected molecular weight fractions for targeted protein quantification using LC-MS/MS
  • Isolate intact proteins to analyze variants, post-translational modifications, alterations
  • Separate protein pull-down components for target protein purification
  • Separation, isolation, and intact recovery of antibodies intact for in-depth characterization
  • Suitable for both bottom-up and top-down proteomics:

Bottom-up proteomics – Broad fractionation for increased coverage. Reduced complexity. Gain molecular weight information to distinguish peptides from multiple gene products.

Top-down proteomics – Intact protein fractionation and isolation. Analyse post-translational modifications, variants and alterations. Native electrophoresis is in development.

GELFREE 8100 Cartridge Specifications

% Polyacrylamide mW Range Optimum mW Range aver. Fraction Width
5% 3.5 – 500 kDa 75 – 500 kDa 40 kDa
8% 3.5 – 150 kDa 35 – 150 kDa 10 kDa
10% 3.5 – 100 kDa 15 – 100 kDa 7 kDa
12% 3.5 – 60 kDa 10 – 50 kDa 4 kDa

GELFREE 8100 Sample Specifications

  • Max sample load volume 150 μl
  • Fixed fraction volume of 150 μl
  • Protein load ≤ 500 μg (≤ 3.3 mg/ml)
  • Conductance ≤ 5 mS/cm
  • pH 8.4 ± 1.0

Request your exclusive demo today!*

Simply enter your details in the above form and we will put you in touch with one of our GELFrEE experts to arrange your demonstration.

*T&Cs: Purchase of GELFrEE Cartridge is required to qualify for a free demo.

 

Simply enter your details below and we will put you in touch with one of our GELFrEE experts to arrange your demonstration.