ACKNOWLEDGEMENT Before getting into the details of the project, i would like to express my wholehearted gratitude to one and

all , who have helped me in doing this project successfully. First of all, i thank my biology teacher Mrs.bindhu and biology lab assistant Mrs. Prema for their support , guidance and their valuable advices. I express my sincere thanks to my group members, Vijay P and Azhar UP, for their valuable contributions for this project. Iam indebted to my parents and sister for thier constant encouragement and support Finally, i would like to thank CBSE for giving me this opportunity to undertake this project. Above all, i thank Almighty for making this venture a success. (next page) Contents (write it) Introduction Electrophoresis is a simple, rapid and highly sensitive tool to analyze proteins.The separation of proteins by electrophoresis is based on the fact that charged molecules will migrate through a gel matrix upon application of an electric field
. Ions run more or less quickly along the substrate according their charge, size, shape, etc. Electrophoresis is widely used to separate substances such as amino acids, proteins, strands of DNA, etc. As in the case of chromatography, people use different supports and solvents according to the substances to be separated and the techniques used (next page) Paper electrophoresis is useful for the separation of small charged molecules such as amino acids and small proteins. A strip of filter paper is moistened with buffer and the ends of the strip are immersed into buffer reservoirs containing the electrodes. The samples are spotted in the center of the paper, high voltage is applied, and the spots migrate according to their charges. After electrophoresis, the separated components can be detected by a variety of staining techniques, depending upon their chemical identity. Electrophoretic techniques have also been adapted to other applications such as the determination of protein isoelectric points. Affinity gels with biospecific properties are used to study binding sites and surface features of proteins. Continuous flow electrophoresis is applied to separations in free solution and has found very useful application in blood cell separation. Recently, High Performance Capillary Electrophoresis (HPCE) has been developed for the separation of many classes of biological molecules (next page)

Paper electrophoresis attained widespread use in the 1950s. It is a scientific method of separating a substance to determine its composition. Wide-ranging ramifications of this method and resulting newer electrophoresis technologies have served scientific and other industries as well.. Scientific Applications The main use of paper electrophoresis is that of analysis. Scientists have used this technique in many scientific experiments as a way to prove or disprove hypotheses or to determine the efficacy of drug treatments. , scientists use a specific paper electr ophoresis test, called the Western blot, to detect the presence of human immunodeficiency virus (HIV) in blood samples. In addition, other blot tests help detect the type and amount of DNA and RNA in sampled materials, which has major implications for fore nsic investigations. Furthermore, paper electrophoresis can be used for testing suitability of municipal water supplies, toxicity of waste water and other environmental components. Drug Testing

Results of these tests help determine the authenticity of suspected counterfeits.: filter paper.000 per square inch of paper. containing a clear fluid in which positively charged part icles of one color and negatively charged particles of a contrasting color reside. (next page) Aim To obtain the blood components . Ink Analysis Inks such as those used to print currency and checks consist of diverse chemical makeups. as well as for signage. polyacrylamidegel. by the technique of paper electrophoresis and also to scrap out the blood smear in the anode to observe under the high power of a microscope. E-Paper E Ink used electrophoretic technology in pioneering electronic paper display (EPD) technology. The low voltage required makes these items tremendously inexpensive to operate. Furthermore. in the anode of electrophoresis apparatus. sports authorities such as the International Olympic Committee have used the technology to screen athletes suspected of ingesting illegal performance -enhancing supplements. When an electric field is applied to a paper or film printed with the ink. an electrical DC power supply two electrodes table salt baking soda saffranin 600ml water (next page) procedure . all the particles of one charge rise to the surface. depending on the polarity of the electricity This technology is being used for watches. Investigators and forensic scientists have used paper electrophoresis since the 1950s to analyze inks. mobile phones and other handheld electronics.e. about 100. (next page) Apparatus required two small basins a support (i. or a capillary tube) . cellulose acetate strips.The drug-testing industry has used paper electrophoresis to determine the presence of illegal or recreational drugs in job applicants and crime suspects. The company invented an electronic ink that consists of millions of minute microcapsules. forgeries and other fraudulent documentation.

draw a line across the filter paper and place a little drop of blood on it . You can obtain this from five. Acidic amino acids will be negatively charged and so they will behave as anions. Place a glass plate on the two basins and on it place a strip of filter paper soaked with the electrolyte. Put an electrode into the electrolyte of each basin and apply 45V in direct current . different little white coloured spots are moved towards the negative electrode. 7. The speed and direction of migration will depend upon the numbers of the characteristic amino acids in the protein's structure. (next page) Observations With the passing of time. Explanation At pH 7 basic amino acids will be positively charged and so they will behave as cations. Similarly at pHs below the isoele ctric point the molecule will have a net positive charge and move towards the cathode. Neutral amino acids will be both positively and negatively charged (zwitterions) and so they will stay in the centre. 9volt batteries connected in series. This strip has to be immersed with each end in the electrolyte in the basin for to complete the electrical circuit . With a pencil. Cover the paper with a second glass plate .(next page) initial procedure 1. The relative charge on each species of amino acid will determine the speed and direction of travel. they will h ave different isoelectric points (next page) Final procedure . As proteins are made of amino acids and amino acids migrate in an electric is observed that. At any pH above the isoelectric point the molecule will have a net negative charge and move towards the anode. Thus they can be separated. place two little basins a few cm (one inch) apart. 4. 2. it depends upon the number of ionisable groups in the molecule. A protein made up of a lot of basic amino acids will not behave in the same way as a protein made up of acidic ones. Pour in the basins an electrolyte made of a teaspoon of table salt and another of baking soda in 300 ml(1 1/2 cups) of tap water . then proteins too will migrate (slowly). Observe the system after two hours 3. Each type of amino acid has its isoelectric point at a particular pH. 5. These amino acids are said to be at their isoelectric point. 6.

remove the electrodes from the basins and take off the cables from the batteries. Biomolecule ²An organic compound present in living organisms.they are of 4 types*Alpha 1 globulins *Alpha 2 globulins *Beta globulins *Gamma globulins 3 Fibrinogen (factor I) is a a soluble plasma glycoprotein. All humans produce these blood components--there are no populational or regional differences. the blood components are observed as greenish blue patches of proteins spread over the red background of RBCs (next page) Result The greenish blue patches observed are made of different protein components of the plasma (serum proteins): globulins (alpha. Take care never to short out the two electrodes. Four of the most important ones are red cells. and plasma. insert a fuse on one of the two cables.2. At the end of the migration of the spots. (next page) KEY TERMS 1. Biological activity ²The specific biochemical function of a biomolecule such as enzyme affinity for a substrate. . 2. beta. gamma). The white spots accumulated at the anode are scrapped with a blade to a slide Add a small drope of saffranin and keep a coverslip over it Observe it. the electrolytes. 4. platelets. . Albumin is essential for maintaining the osmotic pressure needed for proper distribution of body fluids between intravascular compartments and body tissues Globulin is one of the three types of serum proteins. while others are made by the immune system. Some globulins are produced in the liver. These are aggregated together into small patches under the action of electrolytic movement through the strip of filter paper.1. Never touch the electrodes. or the paper strip. 2. synthesised by the liver. albumin and fibrinogen. Do not use high voltages for this experiment. The main protein components of the plasma are: 1 2 Serum albumin. 2. white cells. often referred to simply as albumin is the most abundant plasma protein in mammals. first under low power and then under rhe high power of microscope (next page) Observations Under the high ower of microscope. Blood is a highly specialized tissue composed of many different kinds of components. The fuse will break the circuit when there is too high a current. 3. that is converted by thrombin into fibrin during blood coagulation (next page) Precautions 1. 3.

4. Electrophoresis gel ²A natural or synthetic polymer matrix used as a support for sample separation in electrophoresis.3.jrank. Molecular sieving ²The restriction of molecules migrating through a gel due to their size Bibliography y y y y Icbse. Ehow. Isoelectric point ²The pH at which a molecule has no electric charge and does not move in an electric Wikipedia .

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