Laboratory Exercise #3 Measuring Protein in Solution Abstract The purpose of this lab was to learn about the Biuret assay reaction to determine if it can detect proteins and amino acids; also‚ to understand the process of “salting out” proteins and how to determine the amount of protein in a solution. In order to do so‚ egg white and ammonium sulfate were mixed on ice and then put into the centrifuge. After PBS was added‚ the amount of protein could then be determined. After that‚ 14 test tubes
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In this lab we employed various assays utilizing a biuret reagent‚ coomassie brilliant blue reagent‚ and ultraviolet light in order to determine the identity of six unknown solutions and the concentration of a bovine serum albumin sample. We were given three samples that lacked protein‚ and three samples containing proteins‚ and using a spectrophotometer we assessed the amount of light absorbed versus the light transmitted‚ based on the principles of the Beer-Lambert Law. The three proteins used included
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Purification of Green Fluorescent Protein Introduction: Transformation is used to introduce a gene coding for a foreign protein into bacteria. Hydrophobic Interaction Chromatography (HIC) is used to purify the foreign protein. Protein gel electrophoresis is used to check and analyze the pure protein. Research scientists use Green Fluorescent Protein (GFP) as a master or tag to learn about the biology of individual cells and multicultural organisms. This lab introduces a rapid method to purify
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Daniel Bergey Lab 2: Proteins and Starches Purpose The purpose of lab 2 and both tests with proteins and starches is to determine which substance contains either protein or starch. Hypothesis Proteins: I predict that any substance I test that derives from a living organism is will test positive proteins. Any substance that isn’t from a living organism more than likely will test negative for proteins. Starches: I predict that any substance that contains any level of glucose will test positive
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To purify the protein in the cell lysate from lab 1 through nickel affinity chromatography. Protein purification should result in only one type of protein ideally‚ which is the protein of interest‚ wt-DHFR and mut-DHFR in this case. A pure protein allows for further analysis on the protein to be conducted‚ such as its concentration (Bradford assay)‚ its molecular weight‚ and its biological activity. 2. Overview of experiments Buffer Preparation Add the liquid sodium phosphate‚ solid sodium chloride
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Bita Heydari Lab report 3 The Effects of Differentiation on Enzymatic Activity Introduction HL-60 cells are capable of undergoing differentiation to induce different cell types. HL-60 cells can undergo morphological changes‚ changes in gene expression‚ and changes in protein synthesis. In the past weeks‚ we were able to conclude that HL-60 cells treated with DMSO and HL-60 cells treated with PMA will differentiate into granulocytes and monocytes upon treatment (1). We were also able to observe
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Lab2: Testing for Proteins and Starches In this lab 8 total substances were tested to find out whether they are a Protein or a Starch. It is my belief that only 1 or two of each of the substances in test 1( proteins) and test 2 (starch) will test positive for either protein or starch. For this lab the following materials were needed to complete the experiments in test 1 for proteins: Di water‚ ev milk‚ 50% egg solution‚ 1% sucrose‚ 4 test tubes‚ 1 test tube rate‚ safety glasses‚ pipets and
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If pH > pI‚ then the protein will have a negative charge and if pH < pI‚ the protein will have a positive charge. Buffer I has a pH >5‚ meaning both proteins carry a negative charge and bind to the DEAE (a positively charged resin). (b) pH = pKa + log10(Base/Acid) [Base = mM of sodium acetate; Acid = mM of acetic acid] = 4.7 + log10 (40/40) = 4.7 In order for the catalase to elute from the column‚ it must have lost its negative charge and stopped binding to the DEAE. Lowering the pH
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Abstract There are many methods employed to precipitate proteins out of solution. In this experiment we manipulated many physical and chemical variables in order to achieve purification of a protein via precipitation. In the first part of the experiment we purified the protein casein by modifying it’s pH. In the second part of the experiment we manipulated the ionic strength of albumin in egg whites‚ in a process called salting out. By manipulating these chemical properties we were able to
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Task 1 • Describe the structure of an enzyme as a protein‚ in terms of tertiary/ quaternary structures. 1) Primary Structure This is in reference to the order of way that amino acids are connected to form a protein. These are built up from 20 amino acids‚ and follow these structures o A carbon (the alpha carbon) bonded to the four groups below: o A hydrogen atom (H) o A Carboxyl group (-COOH) o An Amino group (-NH2) o A "variable" group or "R" group 2) Secondary Structure This is in reference
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