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Laboratory Services

Sample Preparation and Fractionation

One of the most important elements of a proteomic experiment is consistent, high-quality sample preparation. Proper sample preparation ensures that protein separation techniques can be successfully performed, that differential analysis will yield meaningful results, and that high quality mass spectrometry data can be readily obtained . Key factors that affect sample quality include the concentration and dynamic range of proteins present in the sample as well as the chemical nature of protein and non-protein components.

NextGen Sciences offers the following sample preparation services upstream of electrophoresis, chromatography and mass spectrometry:

• Preparation of crude protein lysates from tissue and cell pellets
• Differential detergent protein extraction and solublization
• Depletion of abundant proteins from serum (CB or affinity chromatography)
• Chromatographic fractionation
• Enrichment of selected protein classes (affinity fractionation)
• Free flow electrophoresis
• Buffer exchange (dialysis, ultrafiltration)
• Protein concentration (ultrafiltration, gel electrophoresis)
• Solution phase IEF

Gel electrophoresis

Gel electrophoresis is one of the most robust techniques for separating and concentrating proteins from complex biological mixtures. Unlike other protein separation techniques, gel electrophoresis allows for the reproducible comparative analysis and identification of hundreds of proteins in a single experiment. NextGen Sciences offers electrophoretic services both for differential analysis and as a technique for protein concentration and purification upstream of mass spectrometry.

Specific services include:

• Large and mini-format two dimensional gel electrophoresis (2-DE)
• SDS-PAGE
• Visible and fluorescent staining and imaging
• 2-D DIGE
• Western blotting

 

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