A milligram-preparative polyacrylamide-gel electrophoresis apparatus providing reduced eluate dilution

The construction and operation of a preparative polyacrylamide-gel electrophoresis system is described. Emerging protein bands are collected by an intermittent pumping system which is based on the design of Brownstone ((1969) Anal. Biochem. 27, 25–46) . The original pressure-sensitive operation was,...

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Veröffentlicht in:Analytical biochemistry 1976-10, Vol.75 (2), p.363-373
Hauptverfasser: van Jaarsveld, P.P., van der Walt, B.J., le Roux, C.H.
Format: Artikel
Sprache:eng
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Zusammenfassung:The construction and operation of a preparative polyacrylamide-gel electrophoresis system is described. Emerging protein bands are collected by an intermittent pumping system which is based on the design of Brownstone ((1969) Anal. Biochem. 27, 25–46) . The original pressure-sensitive operation was, however, simplified to time-volume operation. Cooling of the gel by a central cooling finger, essentially according to Jovin et al. ((1964) Anal. Biochem. 9, 351–369) , has also been added. To accomodate the polyethylene tubing needed for intermittent collection of protein and also the central cooling finger, it is necessary to polymerize the gel in a mold before it is installed in the gel housing compartment of the electrophoresis cell. Gel concentrations of 5% and higher can be used in this system. Dilution of emerging protein samples by the intermittent collection system is kept to a minimum. This fact, together with simplicity of design makes it suitable for general preparative work with polyacrylamide-gel electrophoresis. Operation of the apparatus and resolution of protein bands are demonstrated by separation of bovine serum albumin polymers and thyroxine-binding proteins in human serum.
ISSN:0003-2697
1096-0309
DOI:10.1016/0003-2697(76)90090-7