Protein Composition in Red Blood Cells in Humans using Polyacrylamide Gel Electrophoresis Name: Emma Claypole Date: Wednesday March 16, 2016 Lab Group: W08, Wednesday morning 2 Abstract The proteins of Bovine red blood cell (RBC) membranes were analyzed using polyacrylamide gel electrophoresis. After analyzing Bovine RBC they were then compared to human RBC counterpart. Following finding the log of each molecular weight of each band, band one showed the highest molecular weight. All five bands viewed were from humans. There are typically 7 bands visible however, in our case only five were visible due to implications within the gel sample. Introduction Proteins are important elements in cellular membranes and give the membranes many of their characteristics. In red blood cells, the meshwork of proteins in and around the membrane gives it strength and flexibility, allowing a cell to squeeze through small capillaries without bursting. Other proteins play roles in transporting material in and out of the cell (Lab Manual, Cell Biology). Polyacrylamide gel electrophoresis (PAGE), with all of its different modifications is probably the most widely-utilized procedure in contemporary biochemistry and molecular biology (Mordacq and Ellington (1994)). In this experiment, we will attempt to determine the molecular weights of the major proteins in the plasma membranes of bovine red blood cells (RBCs). The predictions made are if our protein has similar weights as proteins
This technique separates Rubisco samples based on their size. The electrophoresis has a positive and a negative end. Positive charge proteins are loaded from the positive end and migrate towards the negative end. Negative charge proteins are loaded from the negative end and migrate towards the positive end (Sakthivel & Palani, 2016). The sample that contained the highest molecular weight of Rubisco will travel the shortest distance on the gel while the protein with the smallest molecular weight will travel the longest distance (Sakthivel & Palani, 2016). The size proportion of each Rubisco molecule correlates with the distance traveled. Rubisco will be in its purest form after running through SDS-page since each technique will increase the purity of the protein. If the salting out, the ion exchange and the SDS-page protein isolation techniques are performed on protein Rubisco, then it is purified and separated by solubility, charge, and size. The rationale of this experiment is to isolate the purest form of Rubisco so that it can perform carbon fixation at an optimal
The absorption of proteins as well as thrombus formation on the pH sensors has been known
29. If all the 280 million molecules of hemoglobin contained in RBCs were free in the plasma,
In a cell, diffusion occurs across a selective permeable membrane of the cell that allows the cell to regulate and control the passage of substances. As well as controlling what passes through the membrane, the selective permeability also controls how quickly those allowed substances are able to pass through the cell membrane. When a cell is placed in a hypotonic solution, the substance wants to quickly diffuse through the cellular membrane in attempt to balance the concentration. Due to this, too much of the substance enters the cell causing the cell to swell and rupture, otherwise known as hemolysis. The purpose of this lab is to test if and how the factor of polarity of a molecule affects the diffusion rate of those molecules through a permeable red blood cell membrane in a sample of ovine blood. Testing this factor and its effect on diffusion rate is important in the biological world as it helps understand how our body regulates our own substances and molecules within us, but also how our body will react and respond to other substances such as medicines. It is hypothesized that this factor of polarity will determine how quickly or slowly molecules will diffuse across the cell membrane. Smaller molecules that have relatively low polarity will be able to cross the membrane more easily and at a faster rate than those molecules that
Blood is made up of straw coloured plasma, the matrix, in which various types of blood are carried. Plasma is mainly water where substances are carried such as oxygen and carbon dioxide, nutrients such as glucose and amino acids, salts, enzymes and hormones. Also there is a combination of important proteins which help with blood clotting, transport,
The purpose of this experiment was to determine the effects of tonicity on a cell membrane using red blood cells, potato strips and three unknown solutions (A, B, C). First three slides were prepared containing RBC’s and unknown solutions A, B and C. A control slide was prepared only using RBC’s. After observing each slide under the microscope it was determined that unknown solution A was hypertonic because the RBC appeared to have shrunk. The RBC in unknown solution B appeared to be swollen, therefor, the tonicity of unknown solution B was hypotonic. Unknown solution C showed no change to the RBC shape, it was suggested that unknown solution C was isotonic. To confirm the tonicity
The Journey of a red blood cell begins inside the bone, where the blood cell is made which is the bone marrow. The red blood cells travel around the body within capillaries. Then the deoxygenated red blood cell makes its way to the heart in the vena cava. After the blood cell has made its way through the heart the right atrium (the cell enters the right atrium first) contracts and pushes the blood cell through the tricuspid and into the right ventricle (the parts where the blood cell enters second which is located in the bottom right corner of the heart.) Next the right ventricle again contracts and pushes the blood cell out of the heart through the semi lunar (the section of the heart where the cell enters third). Then finally the deoxygenated
Spectrophotometer; the finding of protein concentration of an unknown sample of BSA, and by using the standard curve.
The alternative method of loading RBCs, involves encapsulating the therapeutics inside the cell. While there are multiple methods for coaxing the cell to uptake a therapeutic, there appears to be a consensus that osmosis driven entrapment is the method that does the least amount of damage to the cell membrane. This method takes advantage of an interesting biochemical feature of red blood cells and can be viewed in figure 2. When RBCs are put in a hypotonic solution
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
Erythrocytes are made in the red bone marrow; and present in the ends of long bones and flat and irregular bones. They pass through several stages of development before entering the blood. There are about 5 million red blood cells in 1 cc of blood.
The membrane of the red blood cell plays many roles that aid in regulating their surface deformability, flexibility, adhesion to other cells and immune recognition. These functions are highly dependent on its composition, which defines its properties. The red blood cell membrane is composed of 3 layers: the glycocalyx on the exterior, which is rich in carbohydrates; the lipid bilayer which contains many transmembrane proteins, besides its lipidic main constituents; and the membrane skeleton, a structural network of proteins located on the inner surface of the lipid bilayer. In human erythrocytes, like in most mammal erythrocytes, half of the membrane mass is represented by proteins and the other half are lipids, namely phospholipids and cholesterol.[29]
Proteins are the basis of the protoplasm (fluid living content of the cell that contains the cytoplasm and cell nucleus) and are found in all living organisms. Proteins make up the bulk of animals body’s non-skeletal structure. As enzymes, they catalyze biochemical reactions; as antibodies, they prevent the effects of invading organisms; and as hormones, they control metabolic processes (C. Bissonnette, 2011). The Biuret test was used to detect the presence of peptide bonds within proteins, and they were found present in test tube #9 (control for peptide bonds).
The other type of artificial blood is more of a blood substitute as it is derived from either outdated bovine or human red blood cells. It is known as Hemoglobin Based Oxygen Carriers (HBOC), Hemoglobin which is the oxygen carrying protein molecule found in red blood cells is extracted from the obsolete red blood cells through ultrafiltration and purification. The Hemoglobin must undergo specific processes in an attempt to prevent the Hemoglobin from disassociating from its natural four-chain configuration (Fridey 3). There is numerous methods of chemically altering the Hemoglobin to increase the molecules size so it does not dissociate and break down. The two main processes of enlarging the
When RBCs are suspended in an isotonic solution, nothing should be observed as there will be no net water movement between the RBCs and the solution. Thus, when 200 µL of blood was added to 10mL isotonic saline (0,154 M NaCl), voltage of 0V is recorded. When RBCs are suspended in a hypertonic solution, for instance, 0.4 M NaCl, RBCs shrunk due to decrease in cell volume as water diffused out of the cells by osmosis. The protein concentration within the cells become greater and more light is scattered and a negative