Off-line Separation

1 dimensional & 2 dimensional gel eletrophoresis (SDS PAGE & 2DE) and in-solution electrophoretic partitioning are important off-line separation methods for large biomolecules such as proteins and nucleic acids. The BMSF has a comprehensive range of separation devices and mass spectrometers to address most analytical problems suited to mass spectrometry.

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GE and BioRad mini and large format SDS-PAGE systems

Proteomic work can involve the use of two-dimensional polyacrylamide gel electrophoresis (2-D PAGE or 2-DE). In 2-DE, "first dimension" electrophoresis in a polyacrylamide gel with a pH gradient and high urea concentration is followed by a "second dimension" separation in an SDS-PAGE gel. SDS-PAGE can also be used independently. Proteins and peptides are separated within the gel according to the % acrylamide and crosslinker, given as %T and %C.

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GE IPGphor/Multiphor Isoelectric Focusing Systems

Isoelectric focusing (IEF) systems are used for: the fractionation of complex biological samples in pathway and systems analysis; quantitation of protein effector molecules; elucidation of protein modifications; and sample clean-up for ongoing analysis. It is used in conjunction with immobilised pH gradients of length 7cm, 13cm, or 18cm for broad range pH3-10, mid range pH4-7, 6-11, or narrow range with 0.01pH unit separating power.

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NuSep ProteomSep/MF10 Electrophoretic Partitioning Systems

This in-solution electrophoretic device is for the partitioning of μl volumes of μg-mg amounts of proteins and or peptides based on surface charge, pH, size in denaturing or native formats. It is typically used to partition contaminant proteins away from lower abundance proteins (eg. Albumin or Ig from other plasma proteins), desalting of biological samples (eg. urine, plasma, tears), enrichment of poorly concentrated and low abundance samples for further analysis. Partitioning down to 1kDa is achievable and is therefore ideal for peptide and small protein work.

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