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Electrophoresis ; 19(12): 2169-74, 1998 Sep.
Article in English | MEDLINE | ID: mdl-9761199

ABSTRACT

Picomolar limits of detection are obtained using the noncovalent, fluorogenic dye, Sypro Red. The size separation of four commonly used sodium dodecyl sulfate-capillary gel electrophoresis (SDS-CGE) molecular weight markers with 8% linear polyacrylamide (PAA) as the sieving matrix is used to construct a calibration curve for molecular weight determinations. SDS-CGE purity and molecular weight determination of purified chorismate mutase-prephenate dehydrogenase (CMPD) from Escherichia coli is shown to be comparable in accuracy with slab gel SDS-polyacrylamide gel electrophoresis (SDS-PAGE). A migration time precision study indicates excellent reproducibility. Sypro red labeling of SDS-bovine serum albumin (SDS-BSA) complexes at nanomolar protein concentrations suggests assay detection limits surpassing those of silver staining. This detectability exceeds that achieved in previous SDS-CGE laser-induced fluorescence studies. This approach is expected to be easily adapted for use with commercial polymer formulations and automated instrumentation.


Subject(s)
Electrophoresis, Capillary/methods , Fluorescent Dyes , Sodium Dodecyl Sulfate , Buffers , Chorismate Mutase/chemistry , Electrophoresis, Polyacrylamide Gel , Escherichia coli/enzymology , Microchemistry , Molecular Weight , Prephenate Dehydrogenase/chemistry , Protein Denaturation , Reproducibility of Results
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