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Electrophoresis

             The separation of native proteins by electrophoresis using agarose gel as a medium.
             The purpose of this experiment was to see which direction certain proteins will migrate according to net charge of protein and pH of buffer used. .
             III. Proteins play an important role in the structure and function of all living things. Amino acids are the protein's building blocks. There are 20 amino acids that are found in proteins. What makes them different is the nature of the R-group attached to the alpha carbon. The amino acids are held together by peptide bonds. These bonds help give each protein it's unique structure and function. These three-dimensional shapes are of critical importance to the protein's function. There are three structural levels that define the three-dimensional shape of a protein. The first of these called the secondary structure are the alpha helices and the beta pleated sheets. These structures are held together by hydrogen bonds. The second called the tertiary structure is brought about by the interactions of the alpha helices and the beta pleated sheets as well as any other folds, turns or straight stretches in the molecule. The hydrophobic parts and the hydrophilic parts assume their posit!.
             ion so as to either be near the water or away from the water respectively. Lastly is the quaternary structure, which deals with the arrangement of the many polypeptide chains in a variety of noncovalent bonds.
             As stated in the beginning, the purpose of this experiment is to use electrophoresis to separate proteins. Electrophoresis is the movement of charged molecules under the influence of an electric field. Since proteins and amino acids are charged molecules, when they are placed into an electric field they will migrate depending on the pH value. The agarose gel has become a regular medium for this type of experiment because it serves as a matrix for the buffer in which the sample molecules will be traveling.


Essays Related to Electrophoresis

1. Electrophoresis

... composed of mainly nucleic acid within a protein coat) and small organelles. Nucleic acid is an organic substance that plays a role in the storage and replication of hereditary information. Electrophoresis" principle is that the charged particles of a sample migrate in an applied electrical field. In conducted in solution (homogeneous mixture of two or more substances), samples are separated according ... their surface net charge density (ratio of mass of a substance to its volume). The most frequent applications, however, use gels (polyacryamide, agarose) as a support medium. Proteins e...

  • Word Count: 1589
  • Approx Pages: 6
  • Grade Level: Undergraduate

2. Electrophoresis

... composed of mainly nucleic acid within a protein coat) and small organelles. Nucleic acid is an organic substance that plays a role in the storage and replication of hereditary information. Electrophoresis" principle is that the charged particles of a sample migrate in an applied electrical field. In conducted in solution (homogeneous mixture of two or more substances), samples are separated according ... their surface net charge density (ratio of mass of a substance to its volume). The most frequent applications, however, use gels (polyacryamide, agarose) as a support medium. Proteins e...

  • Word Count: 1595
  • Approx Pages: 6
  • Grade Level: Undergraduate

3. Gel Electrophoresis

... a process in which nucleic acids or other proteins are separated by way of an electrical current on the basis of size or weight. In gel electrophoresis DNA is cut to fragments by use of a special type of enzyme known as restriction endonuclease (such as EcoR1 and HindIII the enzymes used in our lab) of varying lengths. ... ...

  • Word Count: 397
  • Approx Pages: 2
  • Grade Level: High School

4. Separating Biological Macromolecules by Agarose Gel Electrop

... and trace level determinations. Analysis times are in the region of 1-30 minutes depending on the complexity of the separation. Immunoelectrophoresis: There are two major types of electrophoresis: protein electrophoresis and immunoelectrophoresis. Immunoelectrophoresis is used to assess the blood levels of specific types of proteins called immunoglobulins. In immunoelectrophoresis, a gel is prepared with alternating wells and slots ... , placed orthogonally with respect to the gel, that receive short alternating pulses of current. PFGE allows whole chromosomes and large portions of chro...

  • Word Count: 1328
  • Approx Pages: 5

5. Experiment - Genetically Modified Foods

... in a simple reaction with the proper primers(2). The ability to amplify these DNA sequence allows scientists to further analyze these DNA sequence using gel-electrophoresis. By measuring the length of base pairs, foods can be distinguished from conventional food or GM food. PCR has been found to be appropriate ... ensure the Master Mix is functioning properly. The 6 solutions are placed in a freezer to allow the reaction to go through. To set up for electrophoresis, the gel is placed in the chamber and filled with buffer. Add 10uL of Orange loading dye into each sample. Load 15uL of solu...

  • Word Count: 1122
  • Approx Pages: 4
  • Has Bibliography
  • Grade Level: High School

6. The Cystic Fibrosis Gene

... are known to travel with genetic disorders.3 The RFLP for cystic fibrosis was discovered through the techniques of Somatic Cell Hybridization and through Southern Blot Electrophoresis (gel separation of DNA). By using these techniques, three RFLP's were discovered for CF; Doc RI, J3.11, and Met. Utilizing ... sequence obtained from chromosome jumping. To get these probes, DNA from a horse, a cow, a chicken, and a mouse were separated using Southern Blot electrophoresis. Four probes were found to bind to all of the vertebrate's DNA. This meant that the base pairs within the probes discove...

  • Word Count: 2162
  • Approx Pages: 9

7. Microsatellites In DNA

... 7 minutes for the last extension. This process is not very long, but must be repeated at least 30 times for effective amplification. The newly made DNA is then ran through electrophoresis. This is a process which separates DNA, RNA and proteins by size or charge. The sample was placed in an argarase gel. The sample is then ran through 50-60 V for ... now find out where their blue eyes come from or why they have brown hair. Microsatellites, minisatellites and satellites all cause mutation in gene sequences. Because of the functionality of electrophoresis people can now see how the differ...

  • Word Count: 1018
  • Approx Pages: 4
  • Has Bibliography
  • Grade Level: High School

8. Sickle Cell Disease

... . The severity of sickle cell disease varies from person to person, but most of the time sickle cell can lead to a fatal outcome. Sickle cell disease is diagnosed by hemoglobin electrophoresis. Hemoglobin electrophoresis is a blood test that can detect different types of hemoglobin, including hemoglobin S, the main component of sickle cell disease. Most states routinely screen newborns for sickle cell disease with ... ...

  • Word Count: 1101
  • Approx Pages: 4
  • Has Bibliography
  • Grade Level: High School

9. The Transformation of Bacterial DNA

... . The reason why there is RNA in our samples is because we did not use any RNAse or other enzymes to degrade it. The following week involved the use of electrophoresis to separate sample components and isolated the GFP protein. When preparing the gel the 1% concentrated gel is used because it is easier to handle. We also used a DNA loading buffer ... the supernatant when lysed cells are centrifuged. Under the UV light, my supernatant glows and my partner's did not. In the final lab, we analyzed our extracted proteins. When using electrophoresis, I had two samples, one heated and one non-...

  • Word Count: 811
  • Approx Pages: 3
  • Grade Level: High School

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