Enzyme assay

The assay was performed as described in section 3.2.2.2. P4 concentrations were analysed in spent culture media for cells cultured for 48, 96 and 144 hr in the present of melatonin 0, 20, 200 and 2000 pg/ml under both 5% O2 and air condition. Samples were diluted 100 fold in PBS and analysed in duplicate with standard (0, 0.5, 1, 2, 5, 10 and 20ng/ml), blank and quality control. To avoid variation among plates, all samples were run together. Inter and intra assay % CV was (11.1) and (5.1) respectively. 4.2.3.2 Oestradiol…

Skill #28 Kirstin Krueger #21839116 ELISA is an abbreviation for "enzyme-linked immunosorbent assay." An ELISA test uses different parts of the immune system, different antibodies, and color changes to detect immune responses in the body. CPLi, or Canine Pancreatic Lipase Immunoreactivity, is a specific lipase that is only found in canine pancreatic tissues. The CPLi ELISA snap test is made by Idexx and is used to measure the concentrations of pancreatic lipase in the serum of the patient.…

NADH is at 340nm, but the peak absorbance of NAD+ is 259nm (Powers, et al., 2007). Enzyme assays are performed to measure the rate of an enzyme-catalysed reaction, known as the enzyme activity. Many factors can affect the enzyme activity (including temperature, pH, substrate concentration), so all conditions apart from the one being quantified should be standardised. The…

Infectious Diseases. 15:523. DOI 10.1186/s12879-015- 1248-6. This article compares and contrasts the sensitivity and specification of Enzym-linked Immunosorbent Assay (ELISA) and the Widal test for diagnostic purposes. To compare the two assays the researchers used blood culture collected from febrile patients for the last three years. The paper clime the widely used Widal testing method has a poor sensitivity as well as up to three days of delayed response time. In addition, the Widal testing…

Most proteins are enzymes, which have an essential role in biological catalysis by increasing the rate of a reaction. The experiment conducted included an enzyme assay with the enzyme catalase and the substrate hydrogen peroxide. To complete the assay, the catalase enzyme was added to the hydrogen peroxide buffered solution. Every thirty seconds, portions from the tube was removed and placed into the labeled tubes. Based on the degree in color of each sample, a different absorption value would…

concentration must be known in order for proteins to be used in enzyme reactions, as the correct stoichiometry must be known. Another example is protein purification; during protein purification the quantity of protein present must be known to determine the yield1. The quantification of proteins is determined by methods known as protein assays. There are different protein assays used to determine the concentration of proteins. Three different types of protein assays used by biochemists include…

Oxaloacetate, found in the mitochondrion, is a four-carbon molecule. In this project we performed a Bradford Assay and an SDH (Succinate Dehydrogenase) Assay. The goal of this project was to work with enzymes to gain a better understanding of how enzymes function and how they are impacted with the introduction of a variety of factors. A significant part of the experiment was determining the protein concentration of the prepared mitochondrial fraction which will be further explained. The reason…

Biochemical foundation for each of the four enzymatic assays performed on each cellular fraction. We performed various assays on each of the four log phase culture fractions, as well as the extracellular fraction from a stationary phase culture, to test the localization of the enzymes. For the protease assay, a specific soluble chromogenic substrate called azocasein was employed to determine presence of enzyme activity. The degradation of this substrate in trichloroacetic acid produces a colour…

Assessment of enzyme producing efficiencies of endophytic fungi Extracellular enzymes assay was conducted to investigate the production of enzymes by the endophytic fungi Amylase activity Amylase enzyme was assessed by growing the fungi on glucose yeast extract peptone (GYP) agar medium (glucose- 1g, yeast extract- 0.1g, agar- 15 g, distilled water 1000mL and pH 6) containing 1 % soluble starch after 5 days incubation , the plates with fungal colony were flooded with 1% iodine and 2% potassium…

4. Multiplex Polymerase Chain Reaction Multiplex PCR is molecular biology technique for amplification of multiple targets in a single PCR reaction. Indeed, multiplex PCR is a modification of conventional or realtime PCR in which two or more different PCR products are amplified within a single reaction (Henegariu, 1997). In a multiplexing assay, more than one target sequence can be amplified using multiple primer pairs in a reaction mixture. As an extension to practical use of PCR, this technique…