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 the Bicinchoninic Acid (BCA) protein assay, Modified Lowry protein assay, and the o-Phthalaldehyde (OPA) Fluorescent Protein…
Then, using the same procedure as before, the solutions ' ultraviolet absorptions at 450 and 595 were determined and recorded in Table 4. As for the BCA assay, 100 microliters of each ovalbumin dilution were combined with 900 microliters of BCA, and the resulting solution were analyzed with a spectrophotometer in order to determine the uv absorbances at 560 nanometers (Data displayed in Table…
Homogenates of liver were provided. The homogenates were made by homogenizing liver in a 1:20 w/v (1g: 20 ml) ratio in a sucrose-phosphate buffer, in which 250mM of sucrose and 50mM NaPO, with a pH of 7.4 were added. This information was used to conduct an experiment to determine the concentration of DNA, protein, and carbohydrates in different homogenate samples, including heart, liver, and kidney of Bos taurus, while also looking for the diverse protein sizes found in each homogenate sample.…
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)…
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.…
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…
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…
Trypsin is also known to act on itself resulting in both active and inactive forms of trypsin during storage (Sugumaran). In this experiment, affinity chromatography was used to extrapolate the active form of trypsin from the inactive form of trypsin. A column containing P-amino-bezaidine was used to form a bond with the active form of trypsin in the matrix (Sugumaran). Two assays were performed. The first assay was to determine if there was any active trypsin by employing…
oxygen3. Serratia Marcescens is a gram negative bacterium. Gram negative bacteria have a thin cell of a single layer of peptidoglycan that is enclosed by an outer membrane. The outer membrane is surrounded by special phospholipids composed of fatty acids called lipopolysaccharides that are attached to a glucosamine…
neutralization reaction. A neutralization reaction is a chemical reaction in which an acid and a base react to form an ionic compound (a salt) and water; the resulting pH is closer to 7. The general chemical equation for a neutralization reaction is: Acid + Base Water + Ionic Compound This can also be considered as a double displacement reaction. However, since the reacts are an acid (carbonic acid) and a base (sodium hydroxide) and the products are a salt (sodium carbonate) and water (H2O),…