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…