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New Automated Systems for Size-Fractionation of Protein Samples

Sage Science
9 Sept 2013

This application note describes two new systems for automated preparative electrophoresis of protein samples. It is demonstrated that these systems provide increased reproducibility and ease-of-use for procedures that require gel isolation of proteins, such as top-down proteomics studies and other protein mass spectrometry methods. The BluePippin system is useful for applications where a single targeted fraction containing a protein of interest is needed. The ELF system is useful for broader proteomic studies including top-down proteomics and GeLC-MS.

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BluePippin Size Selection System

Sage Science

The BluePippin is a preparative electrophoresis platform for collection of size-selected DNA or protein samples. The system automates DNA or protein size selection using disposable, pre-cast agarose cassettes to extract fractions according to user-defined software input. Samples are electro-eluted from agarose and collected in buffer. The platform is used for the construction of next-gen sequencing libraries, or upstream of protein mass-spec. For DNA applications, the BluePippin features pulsed-field electrophoresis to collect high molecular weight targets, and can collect fragments (depending on gel type) between 90 bp- 50kb. For protein, the system uses SDS-agarose to collect targets between 18-200kDa. In both cases, a higher quality of sample is extracted, and more reproducibly, when compared to manual gel purification methods.

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ElectrophoresisElectrophoresis equipment is used to separate mixtures of protein, DNA or RNA, based on their electric charge, size and other physical characteristics, by passing them through a medium such as a polyacrylamide gel, an agarose gel or a capillary tube. Electrophoresis equipment includes horizontal and vertical gel electrophoresis chambers, isoelectric focusing systems, 2D electrophoresis and capillary electrophoresis instruments. Precast gels with a gradient can be used or gels can be hand cast. Samples are run alongside ladders or markers to identify the approximate size of a molecule. Separated proteins and nucleic acids in the gel can be stained or probed with fluorescent markers and then imaged and detected using gel documentation instruments, transilluminators, densitometers and scanners. Find the best electrophoresis equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Sample PreparationSample preparation can improve the quality and speed of separation techniques. Products to assist sample preparation include filtration equipment, evaporators, membranes and sieves.Mass SpectrometryMass spectrometry (MS) is a powerful analytical technique used to identify and quantify molecules based on the mass-to-charge ratio of gas-phase ions. It provides detailed information about the structure, composition, and properties of compounds and is widely used across fields such as environmental monitoring, materials science, drug discovery and development, food and beverage testing, and wider chemical research. Key MS techniques include tandem mass spectrometry (MS/MS), liquid chromatography–mass spectrometry (LS-MS) and inductively coupled plasma (ICP-MS). Choosing from these wide range of techniques and technologies can be a daunting task, so keep up to date with scientific applications, performance expectations, and customer reviews here all in one place. Visit our product directory to receive quotes direct from the manufacturer. ProteomicsProteomics is the systemic bioinformatics study of proteins and amino acids, including their structure, size, function and identification. Tools used in proteomics include chromatography, blotting and gels, protein arrays, mass spectrometry and ELISA and associated analysis software. Analyzers and proteomic systems should be sensitive, high resolution, fast and may be automated for high-throughput.Protein PurificationProtein purification is a vital step in drug discovery, therapeutics, biotech and life science research. The purification process typically involves subcellular or membrane protein extraction with cell lysis kits, separation of proteins from cell debris by filtration or spin columns, and the isolation of proteins of interest from other proteins and impurities with affinity purification (including fusion protein tags and antibody binding proteins A, G and L), immunoprecipitation or chromatographic methods, such as ion exchange, size exclusion and immobilized metal affinity chromatography. All purification methods come in multiple formats for your laboratory needs, including agarose or magnetic beads, resins, columns and filter plates. Find the best protein purification equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Size Selection

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New Automated Systems for Size-Fractionation of Protein Samples