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12‐Channel P eltier array temperature control unit for single molecule enzymology studies using capillary electrophoresis
Author(s) -
Craig Douglas B.,
Reinfelds Gundars,
Henderson Anna
Publication year - 2014
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201300526
Subject(s) - capillary electrophoresis , arrhenius equation , atmospheric temperature range , analytical chemistry (journal) , temperature control , chemistry , arrhenius plot , capillary action , molecule , electrophoresis , range (aeronautics) , activation energy , materials science , chromatography , thermodynamics , organic chemistry , physics , composite material
Capillary electrophoresis has been used to demonstrate that individual molecules of a given enzyme support different catalytic rates. In order to determine how rate varies with temperature, and determine activation energies for individual β‐galactosidase molecules, a 12‐channel P eltier array temperature control device was constructed where the temperature of each cell was separately controlled. This array was used to control the temperature of the central 30 cm of a 50 cm long capillary, producing a temperature gradient along its length. Continuous flow single β‐galactosidase molecule assays were performed allowing measurement of the catalytic rates at different temperatures. Arrhenius plots were produced and the distribution of activation energies for individual β‐galactosidase molecules was found to be 56 ± 10 kJ/mol with a range of 34–72 kJ/mol.