Enzyme assay

mitochondria will be low. Succinate dehydrogenase (SDH) is an enzyme located in the mitochondria membrane. FAD is present, it then turns the succinate to cause cellular respiration. SDH will replace with DCIP, which is an artificial electron acceptor. DCIP is used to measure mitochondrial activity in the spectrometer. DCIP is a redox indicator, causing it…

make sure it stays below 20 C. This was done until the solution was homogeneous. The solution was then transferred to a centrifuge tube and centrifuged. The supernatant was aliquoted and stored in a -20 C for use in future experiments. Enzymatic Assay of LDH by Spectrophotometry. One tube of tissue extract was thawed out on ice and a…

1. What enzyme in the infant is lacking? Explain your answer The lacking enzyme in the infant is glucose-6 phosphatase. When blood glucose is low, glycogen breakdown in liver produce glucose-6-phosphate. G6P release glucose into blood using glucose-6-phosphatase enzyme. This patient is lacking this enzyme so the blood glucose level is low. 2. What is the metabolic reason for elevated levels of blood lactate in this patient? What are the biochemical sources of lactate? Because of the deficiency…

P450-GloTM bioluminescent CYP assay Three concentrations of PTV (1, 10, 100 µM) were conducted to detect the luciferin PPXE activity against recombinant CYP3A4 and CYP3A5. A significant decrease in the luminescene activity in both recombinant CYP3A4 and CYP3A5 was observed in 10 µM and 100 µM of PTV (Fig 1). It seems that PTV is a candidate drug for metabolism against CYP3A4 and CYP3A5 enzymes. Metabolite profiling and Identification After incubating PTV with HLMs in the presence of a NADPH…

to help maintain a biological system at the proper pH. A laboratory buffer should be inert in the system being studied. For example, Tris buffer is unsuitable for some protein assays because it reacts with the assay components. Phosphate buffers contribute phosphate ions to a solution, which inhibit some types of enzyme reactions such as alkaline phosphatase. Tris-Borate-EDTA (TBE) buffer and Tris-Acetate-EDTA (TAE) buffers are most commonly used for DNA gel electrophoresis. However, because…

basics of enzymes and how they work. We were able to perform a quantitative assay of the activity of an enzyme in a tissue extract using a spectrophotometer. Also, we had to organize the data that was provided into tables and graphs so we could have the ability to test a few hypotheses. For example, we tested the whether the rate of the reaction was influenced by enzyme concentration, whether the activity of the enzyme was influenced by temperature and also too see if the activity of the enzyme…

Analysis of soil chemical and biological properties Soil chemical analysis and enzymes like urease, protease and nitrite reductase were carried out in the soil analysis laboratory, Guangxi University, Nanning, Guangxi Province, China. Soil pH analysis (soil:water = 1:1) performed by pH meter and Soil organic carbon was determined by dichromate oxidation (Walkley and Black, 1934). Total N in soil was determined by Semimicro-Kjeldahl method (Bremmer and Mulvaney, 1982). Nitrate nitrogen (NO3−-N)…

monophosphate (present in fresh fish samples) degrades (by microbial action or the action of other endogenous enzymes) to inosine, then to hypoxanthine and xanthine. Elevated levels of hypoxanthine and xanthine are a sign of spoilage [16]. Thus, developing a stable, sensitive, and selective of hypoxanthine/xanthine sensor may also has applications in food quality control. Widely employed assays for xanthine oxido reductase activity are spectrophotometric quantitation of uric acid, the final…

bioluminescence comes from one of the products of a reaction catalysed by the enzyme luciferase (see biochemistry and metabolism). Vibrio Fischeri are easy to grow and they display chemotaxic behaviour. It is best to grow Vibrio Fischeri at 28 degrees Celsius. (DeLoney-Marino CR, 2013) Vibrio Fischeri is a heterotroph and has the ability to be anaerobic and aerobic which was determined through genome sequencing and chemical assays. One of the ways in which it is able to grow anaerobically is by…

ABSTRACT To examine the enzyme kinetics of chymotrypsin during the catalysis of the synthetic substrate p-nitrophenyl acetate (NPA). Initially, observe the magnitude of the initial burst with regards to the enzyme concentration. Secondly, the analysis of the steady state deacylation rate with regards to the pH of the reaction mixture. Chymotrypsin catalysis is a process that proceeds with two steps, an initial fast step involving the covalent modification of the enzyme to the substrate and the…