Bicinchoninic acid assay

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…

This could be net charge, hydrophobicity, density and solubility of the protein (Reed, et al., 2016). Possible dyes that could be used are coomassie brilliant blue and fluoresamine, (Crennell & Todd, 2017). The Bradford assay also makes use of the amino acid residues that make up the primary structure of the protein being studied, in this case aromatic amino acids as well as the amino acid arginine. It tis these residues that directly bind to the Coomassie dye. Having bound to the protein, the…

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 acyl-biotinyl exchange assay (ABE assay) is a powerful technique that enables a more sensitive, rapid, less time consuming, more qualitative method of observing protein palmitoylation while allowing to detect small changes There are four basic steps to this process: immunoprecipitation, HAM biotin labeling, Biotin-BMCC treatment: labelling, and purification of the thiol-biotinylated proteins. Prior to ABE assay, a cell culture must be prepared, and proteins must be extracted from the…

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)…

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…

The data show that in separating the sample mix using size-exclusion chromatography, or gel filtration chromatography, the hemoglobin was able to be separated from the vitamin B12 suggesting that the data are consistent with the hypothesis. This is evident in the data since the concentration of protein decreased with each collected fraction after the first fraction that collected the protein, fraction 2, and leveled off. This is shown by the decreasing absorption measurements in Table 3. The…

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…