Fachbereich Instrumentelle Analytische Chemie / Bio- und Polymeranalytik
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Electrophoresis & Biosample Preparation

 

Bioanalyzer

The 2100 Bioanalyzer system (Agilent, Waldbronn, Germany) was initially developed for electrophoretic separation of proteins, DNA, and RNA in the chip format by application of fluorescent detection.

Analyte separation is based on a sieving matrix (linear polymer) pressurized into photolithographically etched channels of corresponding chips prepared from soda lime glass (Caliper Technologies Corp., Mountain View, CA). Following electrophoretic injection of samples to the separation channel, analytes are separated in an applied electric field in the presence of SDS by their ability to pass through the applied sieving matrix (capillary gel electrophoresis in analogy to SDS – PAGE analyses). Following separation, analytes are detected via a red laser (maximum excitation / emission wavelength at 635 / 685 nm, respectively).

In addition to applications mentioned above, recent developments included a setup to allow for separation of proteins with a molecular weight exceeding 200 kDa (which in standard electrophoresis can only be separated by application of elaborate agarose gels) and electrophoretic separations of particles in the absence of any sieving material to allow for the analysis of larger assemblies like viruses, liposomes and bionanoparticles.

Thin Layer Chromatography (TLC)

Thin layer chromatography is a commonly used technique for the analysis of sample compositions without high instrumental effort. Besides various sample compositions, the method has been established in our lab for the simple and though comprehensive separation of lipid samples based on their polarity and fatty acid side chain length.