Total Serum Protein Determination Lab Report

The homogenates provided were made by homogenizing tissues in a sucrose phosphate buffer in a 1:20 ratio. The protein concentration in bovine cells was measured by diluting the homogenate with a 1:5 ratio; 50 microliters of homogenate and 200 microliters of water. Then 5 known protein concentration samples which were 0.4, 0.8, 1.2, 1.6, 2.0 mg/ml of bovine serum were used to determine absorbance with a spectrophotometer. Two additional samples were made; one was blank and the other was for the specific homogenate sample. Then 3 microliters of bradford assay reagent, which indicates the amount of protein present

The specificity of albumin binding experiment was to determine the binding interactions that occur between serum albumin and three synthetic dyes with the use of electrophoretic procedure. Whole blood, or plasma. Clots upon standing and if the clot is removed, the remaining straw colored fluid is called serum. The major protein in serum is albumin which functions as a carrier molecule for the transport of certain small molecular weight compounds in blood. Molecules that bind to serum albumin are fatty acids, hormones and some synthetic dyes. In this experiment the synthetic dyes used are Bromophenol Blue, Ponceau S and Orange G. we observed that free dyes not bound to albumin migrate faster that albumin or dyes bound to albumin. This

Protein Assay: The Pierce BCA Protein Assay (Thermo Scientific) is a detergent-compatible formulation based on bicinchoninic acid (BCA) for the colorimetric detection and quantitation of total protein concentration. A series of standard solution of Bovine Serum Albumin (BSA) ranging from 0-2000 µg/ml was prepared from a stock solution of 2 mg/ml BSA. 25ul of diluted crude (1:500, 1:250), desalted (1:100, 1:50), and 6 peak fractions from cibarcon blue column (1:10, 1:5) were loaded in microplate along with 175ul of BCA working reagent. Microplate was incubated for 30min at 370C and then the absorbance was measured at 562nm.

After weighing dialysis tubing of starch/sodium sulfate and adding the solution to two test tubes, the tubing was placed in a beaker containing a solution of albumin and glucose. Next, 1.0 mL of albumin and glucose were then placed in two test tubes labeled solution start. The tubing in the albumin/glucose solution was kept inside the solution for 75 minutes. Every 15 minutes the solution and tube was mixed (Keith et al., 2010).

Enzymes are catalysts that function to speed up reactions; for example, the enzyme sucrose speeds up the hydrolysis of sucrose, which breaks down into glucose and fructose. They speed up reactions but are not consumed by the reaction that is taking place. The most important of the enzyme is the shape as it determines which type of reaction the enzyme speeds up. Enzymes work by passing/lowering and energy barrier and in doing so; they need to bind to substrates via the active. Once they do, the reaction speeds up so much more quickly than it would without the enzyme. Coenzymes and cofactors aid the enzyme when it comes to binding with the substrate. They change the shape of the active site so the substrate can bind properly and perform its function.

Colorimetric assay is a process of determining a concentration of a solution based on absorbance of light. The purpose of this lab is to determine if the Bradford assay is an accurate way to determine an unknown concentration of two samples of protein. The Bradford assay is done by measuring wavelength of light passing through a cuvette filled with Bradford dye and concentrations of PBS and proteins. After the cuvettes are mixed they are placed into a spectrophotometer to measure wavelength. The wavelength given will be used to plot a standard curve based on concentration (x-axis) and wavelength (y-axis). The standard curve is then used to measure an educated guess on the concentrations of unknown protein concentrations. We hypothesized that if we use the Bradford assay and colorimetric spectrophotometry we can determine an accurate concentration of two unknown concentrations of proteins. The results of this lab failed to reject our hypothesis based on accurate measurements of protein concentrations. The standard curves are drawn with a linear increasing slope. The Bradford assay is an accurate way to demine the concentration of an unknown concentration.

In week one, only one unknown was found to contain protein. One of the unknowns in the first experiment actually produced a green-blue participate and that was unexpected because that did not show up in our control group. The purpose of the experiment conducted in the second week was to determine the concentration of starch in the supplements. There was a slight stray in this experiment. The 60µg/mL concentration of the control noticeably falls off the line of best fit more than any other point. The unknown products tested contained anywhere from 19-55µg/mL of starch. A way to improve this experiment in the future is to find the concentration of the sugar and protein along with the starch. This way, one would be able to see if there was a relationship between

A cell, the building block of all living organisms, is composed of four fundamental biomolecules: proteins, carbohydrates, sugars and lipids. Proteins provide a vast amount of functions cells such as they serve as enzymes, provide structural support to cells, and act as antibodies. Reagents are used to spark a chemical reaction. The reagent used to detect protein traces in a substance is Biuret’s. Biuret’s will turn purple if proteins are present and blue if they are none. Biuret’s copper particles, have a charge of +2, are diminished to a charge of +1 when peptide bonds, which are in proteins, are present, creating the color change. Polysaccharides, which are carbohydrates, are most notably known to provide energy to the body, but they also help in breaking down fatty acids. Iodine is the reagent used to determine whether a substance has starch in it. The iodine/starch complex has energy levels that are only for retaining unmistakable light, giving the complex its extraordinarily dark black-blue shade. If there is no starch found, iodine will remain its natural yellowish-brownish color, but if starch is present, iodine will turn blue-black. Monosaccharides, which are sugars, like polysaccharides, provide the body with energy. To detect monosaccharides, the reagent, Benedict’s, is used. Benedict’s reagent is added to a test tube, then it is placed in

Chemistry is composed of many molecules and changes of properties in the matter of science. The elements on the periodic table arranged from metal to non metals, plays an important role when it comes to making a compound and mixture. Each of the element have their own unique atomic mass which matters when it comes to organizing the periodic table and when there is a calculation. There are lots of importance about the atomic mass of each element that will be discussed later in this essay. An experiment performed using the mass of zinc and copper has determined whether which element have excess remaining. Will there be a difference if an alkali metal was used?

The enzyme-linked immunosorbent assay (ELISA) is a common laboratory technique used to measure the concentration of an analyte (usually antibodies or antigens) in solution. In the practical anti-BSA antibodies that had undergone serial dilutions were added to a BSA solution in an ELISA plate with goal of seeing how the concentration of anti-BSA antibodies would affect the colour change of the BSA solution. The results clearly showed a direct correlation as the more diluted the anti-BSA antibody solutions became the lower the Wavelength readings at 405nm, which showed that there was less of a colour change.

Albumin levels that below (3g / dL) in hepatitis should be raise suspicion of the chronic liver disease such as cirrhosis of the liver, which reflects to decreased synthesis of albumin.

In the creatinine colorimetric assay the absorbance of the yellow creatinine-picric complex, which is formed from the reaction between creatinine and picric acid, is read spectrophotometrically. Using spectrophotometric data one can determine the relationship between creatinine concentration and absorbance by fitting an equation to the creatinine and absorbance data. The equation sets a standard which can be used to determine the concentration of creatinine in solutions, such as blood and urine, which contain creatinine.

The primary function of the digestive system is to transfer nutrients, water, and electrolytes from the food consume into the body’s internal environment. The ingested food is essential as an energy source, or fuel, from which the cells can generate ATP to carry out their particular energy-dependent activities such as contraction, transport, synthesis, secretion and even renewal of body tissues. Three primary categories of food ingested by humans which are carbohydrates, proteins and fats emerge as large molecules. These large molecules cannot cross plasma membranes intact to be absorbed from the lumen of the digestive tract into the blood or lymph; hence, it must undergo degradation in size (Sherwood, 2013). This

Name ____________________________ I) Introduction All cells contain four major types of macromolecules: carbohydrates, lipids, nucleic acids, and proteins. In today’s lab, we will be studying three of the four-proteins, carbohydrates and lipids. Various chemical tests can be used to detect the presence of each of these molecules. Most of the tests involve a color change visible to the eye. If a color change is observed, the test is considered positive. If the color change is not observed, the test is negative, indicating that a particular molecule is not present. In all the chemical tests we will be performing, we will also be using a control. In most cases, the control will be a sample of

Carbohydrates are the product that made up from carbon, hydrogen and oxygen. Carbohydrates are form by the combination of carbon dioxide and water molecules. The carbohydrates contain two specific functional group in it which is the hydroxyl groups and carbonyl groups.A reducing sugar is a type of sugar with is an aldehyde group.This means that sugar can act as a reducing agent.The procces of reducing sugar is isomerisation,example of reducing sugar islactose,maltose,glucose and fructose.All monosaccharides are capable of reducing other chemicals such as copper (II) sulphate to copper oxide.Beside that disaccharides such as maltose and lactose are reducing sugar,however sucrose is non reducing