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Catalytic assays are performed to determine the amount of the enzyme or substrate in a sample quantitatively. Also, it can be used to prove presence or absence of the enzyme in a distinct specimen, like an organism or a tissue, as a qualitative approach. It measures parameters that indicates the efficiency of the biocatalyst, such as KM, kcat and Vmax. Numerous procedures for catalytic assays of different biocatalysts are already documented or cited in various reference books, but optimization is still necessary, since even accurate observance gives no guarantee of an unequivocal outcome. Effort is usually required to determine the optimum condition of a biocatalyst.
Even though catalytic assays are quite common in academic research and industrial applications, it is hard to establish a general standardization of them, due to the divergent features of enzymes. To achieve the goal, many factors usually need to be modified: reaction pH, buffers and ionic strength, temperature, concentrations of substrates and cofactors, concentration of the biocatalyst, concentrations of additives such as protease inhibitors, and other specific conditions. For high-throughput assays, techniques are developed based on labelled substrates or sensing the reactions of unmodified substrates. Applications with fluorescent probes or mass spectrometry (MS) are typical examples. Recently, microfluidic devices have become an important tool for enzyme assays in more general systems, as they offer the potential to screen for a wide range of applications.
Creative Enzymes offers the service of catalytic assays for the best evaluation of target biocatalysts. We have extensive experience with various types of biocatalysts and therefore offer professional one-stop services:
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