MLAB 2401 Chemistry
Laboratory:
Electrophoresis
Laboratory: Electrophoresis
Points= 5
Objectives:
1.
Summarize the principle of electrophoresis.
2.
Identify normal peaks on electrophoresis protein bands.
3.
Correlate electrophoresis patterns to disease conditions.
Principle:
Plasma contains over one hundred individual proteins, each with a specific set of functions
and subject to specific variations in concentration under different physiologic and
pathogenic conditions. They interact with virtually all body tissues or cells and they are
intimately related to protein metabolism in the liver.
Plasma proteins perform a great many physiologic functions. They serve as transport
molecules for lipids, hormones, vitamins, and metals. They help maintain osmotic balance
and serve as enzymes, complement components protease inhibitors, or kinin precursors.
They play an important role in hemostasis (as clotting factors), the regulation of cellular
activity and function, and in the defense against infection (immunoglobulins). They
contribute to the nitrogen needs of the body.
The key roles which plasma proteins play in bodily function, together with the relative ease
of assaying them, makes their determination a valuable diagnostic tool as well as a way to
monitor clinical progress.
Serum proteins have been separated into major fractions or groups for quite some time
using solubility principles (“salting out” and ultracentrifugation techniques separating
fractions (4.5s, 7s and 19s) based on molecular weight.
The principle of electrophoresis is based on the fact that a charged particle placed in an
electrical field will migrate toward one of the electrodes of the field depending on the:
electrical charge on the particle
size of the particle
strength of the electrical field
nature of the medium used to support the particle during the migration
process
 solubility properties
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Page 1 of 5
MLAB 2401 Chemistry
Laboratory:
Electrophoresis
Reagents, Supplies, and Equipment:
1.
Computer
2.
Gel electrophoresis animation website:
http://learn.genetics.utah.edu/content/labs/gel/
Procedure:
1. Instructor-Lead activity
a. Instructor will show students various electrophoresis patterns.
b. The student will notate possible disease state on result form.
2. Individual activity
a. Navigate your computer to:
http://learn.genetics.utah.edu/content/labs/gel/.
b. Press the forward button by left-clicking on your mouse.
c. Read the narration on the screen and answer appropriate questions found
in the study questions area.
3. Submit to report form.
References:
1. Gel Electrophoresis Virtual Lab. (2012). Retrieved from
http://learn.genetics.utah.edu/content/labs/gel/
Page 2 of 5
MLAB 2401 Chemistry
Laboratory:
Electrophoresis
Laboratory: Electrophoresis
Result Form
Points=4
Name:_____________________
Date:______________________
Electrophoresis Pattern Interpretations
Specimen
Student Result
Instructor result
#D
#E
#G
#H
Page 3 of 5
MLAB 2401 Chemistry
Laboratory:
Electrophoresis
Laboratory: Electrophoresis
Study Questions
15.5 points
Student Name:_________________
Date:________________________
Instructions: Using the tutorial and the Electrophoresis PowerPoint presentation, answer
the below questions. All questions worth one point unless notated.
1 pt.
1. What is a gel?
1pt.
2. What is the purpose of the electrical current in an electrophoresis system?
1pt.
3. What do bands in the gel or supporting medium represent?
1pt.
4. Describe the steps in electrophoresis.
1pts. 5. What are the functions of the buffer?
1pt.
6. In gel electrophoresis, what is the purpose of the DNA size standard?
1pt.
7. In gel electrophoresis, what is the proof that a current is running through the gel?
1pt.
8. What result would the student expect to see with smaller protein molecules?
Page 4 of 5
MLAB 2401 Chemistry
Laboratory:
Electrophoresis
1pt.
9. What is the charge of a cathode? Anode?
1pt.
10. Which ions travel toward the anode? The cathode?
1pt.
11. What property of proteins in the buffer makes electrophoresis possible?
2pts. 12. List four (4) factors that affect migration rates.
2.5pts. 14. Identify the five (5) major protein classes, listing them in order of migration from
the anode to the cathode.
Page 5 of 5