By D.B. Roodyn (Eds.)
Within the face of swift advancements in computerized thoughts, this ebook provides a very good consultant to provide tendencies. It presents info on: rules and terminology of enzyme automation; automated tools illustrated by way of the Technicon process; semi-automatic tools; interrupted-flow and discrete-sampling platforms; single-enzyme research; multiple-enzyme research (M.E.A.); enzyme characterization; calculation of enzyme actions from tool readings; generalized structures for enzyme automation; Appendices I. released computerized enzyme assays, II. Terminology utilized in enzyme automation, III. gear utilized in enzyme automation, IV. desktop software for generalized enzyme automatic approach.
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Additional info for Automated Enzyme Assays
THE TECHNICON SYSTEM 43 to a minimum. However, the system shown in fig. 1 is a useful one, since it may be used as a generalized manifold for many enzyme assays as discussed in detail in ch. 9. Enzyme is introduced through line 5 and the reaction starts as the enzyme mixes with the reaction mixture. This occurs in a jacketed mixing coil (MC,) which is at the temperature of the heating bath, making calculation of the time of incubation somewhat easier. The reaction mixture containing enzyme now enters the heating bath where the bulk of the reaction takes place.
6. Distortion of sampling peak due to Michaelis effect. (a) Typical sampling peak for rise and fall in substrate Concentration, as in fig. 5a. (b) Hypothetical plot of enzyme activity against substrate concentration (Km = 4 arbitrary units, Vmsx = 10 arbitrary units). (c) Sampling peaks expected with (b). ) Subject index P. 218 54 AUTOMATED ENZYME ASSAYS enable one to calculate curve (b) and hence the Michaelis constant and maximum velocity of the reaction. It is clear, therefore, that precise study of the shape of the sample peak could well yield fruitful results.
Fluid containing protein and low molecular weight material is passed into the mixing chamber where it is agitated with a stream of precipitant, such as 5% trichloroacetic acid. The mixture then drips onto a moving strip of filter paper and some of the filtrate (F) is returned to the analyzer. The filter differs from the dialyzer in that the concentration of low-molecular weight substances is the same on both sides of the filter. In calculation of enzyme rates, therefore (ch. 8), one has merely to allow for the dilution caused by the addition of the precipitating Subiecl index D.
Automated Enzyme Assays by D.B. Roodyn (Eds.)