Protein Fractionation : Purification : Isolation : Electrophoresis : Sample Preparation - Gelfree 8100 - Gelfree 8100

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	The Gelfree 8100 Fractionation System is the first product to partition complex protein mixtures into user-selectable liquid-phase molecular weight fractions. Features Intact protein molecular weight fractionation, isolation, and purification Liquid-phase recovery without band or spot cutting Broad mass range fractionation >300 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, pI High loading capacity (>5X more than a 1D gel) Proteins are recovered intact, for complete characterization Applications Simplify and reduce the dynamic range of complex protein mixtures for bottom-up discovery proteomics using LC-MS/MS Fractionate and recover proteins intact for top-down proteomics Isolate and enrich user-selected molecular weight fractions for targeted protein qualtification 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 Components The Gelfree 8100 Fractionation System consists of the Gelfree 8100 Fractionation Station and ready-to-use Gelfree 8100 Cartridges. Gelfree 8100 Fractionation Station and Cartridge The Fractionation Station 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 user's requirements. During operation, the touch screen interface provides feedback on the status and current/voltage that is applied for each channel. Gelfree 8100 Fractionation System Touch Screen Interface The Cartridge Kits Ready-to-use Gelfree 8100 Cartridge Kits complement the Gelfree 8100 Fractionation Station. Each Cartridge Kit contains pre-formulated Running and Sample Buffers, as well as the Fractionation Cartridge. The Gelfree 8100 Fractionation Cartridge contains eight independent channels for molecular weight fractionation and liquid phase recovery. All eight channels may be used simultaneously, or one or more channels may be used, saving the unused channels for use at a later time. Three separate Gelfree 8100 Cartridge Kits are available for protein isolation, purification and fractionation across distinct mass ranges. The Low Mass Cartridge Kit is designed for separation in the mass range 3.5 - 60 kDa. The following image shows an aliquot of each of 12 Gelfree fractions from a yeast lysate partitioned using the Low Mass Cartridge Kit and run out on a 1D gel for visualization. The recommended protocol for the Low Mass Cartridge can be found in our Literature section here. Fractionation of 500 µg Yeast Lysate using the Gelfree 8100 Low Mass Cartridge Kit The Mid Mass Cartridge Kit is designed for separation in the mass range 3.5 - 150 kDa, with resolution between 30 kDa and 150 kDa. The following image shows an aliquot of each of 12 Gelfree fractions from a yeast lysate partitioned using the Mid Mass Cartridge Kit and run out on a 1D gel for visualization. The recommended protocol for the Mid Mass Cartridge can be found in our Literature section here. Fractionation of 500 µg Yeast Lysate using the Gelfree 8100 Mid Mass Cartridge Kit The High Mass Cartridge Kit is designed for separation in the mass range 3.5 - 300 kDa, with resolution between 60 kDa and 300 kDa. The following image shows an aliquot of each of 12 Gelfree fractions from a yeast lysate partitioned using the High Mass Cartridge Kit and run out on a 1D gel for visualization. The recommended protocol for the Mid Mass Cartridge can be found in our Literature section here. Fractionation of 200 µg Yeast Lysate using the Gelfree 8100 High Mass Cartridge Kit All three Gelfree Cartridge Kits may be ordered here or by calling 866.670.9038. Installation Specifications			How it Works Gelfree technology is "Gel-eluted Liquid Fraction Entrapment Electrophoresis. The Gelfree 8100 Fractionation System provides user-selectable, molecular weight fractionation with liquid phase recovery. 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. Performance The Gelfree 8100 system permits effective, programmable fractionation, isolation, and purification across a broad mass range. Combined with its high load capacity and high, liquid-phase recovery , the system provides the highest sensitivity of any pre-fractionation technology. The Gelfree may be operated in fractionation mode, in which the entire sample is partitioned into ≤ 29 selectable MW fractions, or in targeted isolation/purification mode, allowing for extraction of analytes in one or more pre-determined mass ranges. Broad Mass Range Fractionation The Gelfree 8100 Fractionation System offers molecular weight-based fraction targeting over a broad mass range. Using the Low Mass Cartridge Kit, the system is capable of partitioning complex protein samples across the mass range 3.5 - 60 kDa. Using the Mid Mass Cartridge Kit, the system provides fractionation across the mass range 30 - 150 kDa. And using the High Mass Cartridge, proteins are fractionated across the mass range 60 - >300 kDa. Recovery across all mass ranges is ≥ 80%. Intact protein molecular weight-based fractionation with liquid phase recovery: Each of 16 fractions collected from a single channel of the Gelfree 8100 Fractionation System was run on a 1D tris-glycine gel and silver stained. The gel shows the molecular weight based fractions from bovine liver homogenate. Recovery and Reproducibility The Gelfree 8100 Fractionation System uses specialized materials to optimize protein recovery and prevent non-specific loss. In addition, each gel column used in the Gelfree system is comprised of a proprietary composite gel matrix that is robotically cast using state-of-the-art automation. Every gel tube is computer imaged to ensure compliance with strict quality control specifications and each cartridge is barcoded for tracking. This level of quality control and precision-manufacturing uniquely permits the absolute elution time reproducibility provided by the Gelfree system. Gelfree Recovery. BSA was loaded into eight channels of a single Gelfree cartridge and collected over a 7 minute fraction collection interval. Gelfree fractions were run on a 1D gel to visualize the results. Recovery from Gelfree is shown to be >80% and reproducibility of recovery is measured at CV < 10%. Gelfree Reproducibility of Fractionation. 200 µg yeast lysate prepared using the Gelfree 8100 Fractionation System. Fractions 1, 5, and 9 from four lanes across 2 cartridges were run on a 1D gel to demonstrate the gel-to-gel and cartridge-to-cartridge reproducibility of the system. Programmable Methods for Targeted Isolation The Gelfree 8100 Fractionation System is the first product to provide user-programmable molecular weight-based protein purification with liquid phase recovery. Simple adjustment of the fraction collection intervals using the touch screen interface allows isolation of analytes at a targeted molecular weight in a single fraction with near quantitative recovery. To develop a method for targeted protein isolation using the Gelfree system, the user samples the elution window at high frequency over the expected mass range. The peak width is plotted and the fraction collection interval defined. For all subsequent runs, the optimized fraction collection interval is used to isolate and purify the target analyte in a single fraction. Fraction collection windows for a given analyte depend on the amount of total protein loaded. Programmable Protein Isolation at Varying Protein Load. Bovine serum albumin loaded into the Mid Mass Gelfree 8100 cartridge and collected per standard procedures. Peak width varies between 2 and 7 minutes, FWHM. Width increases at higher loading. Fractions were collected at 90 second intervals to define the elution window. Using this elution time data, collection times were optimized for maximum recovery. The gel image on the right shows the Gelfree fraction (GF) containing albumin, 1 µg and 25 µg, compared to control. More Information For more information, watch the video, which shows step-by-step operation of the Gelfree Fractionation System, or contact an application scientist to discuss your specific application.