Science Fair Proposal
Whey Protein Electrophoresis
Norman, Ashton, Edwin
Title: Use of Electrophoresis to determine different compounds within whey protein
Question: Can polyacrylamide gel electrophoresis separate different proteins from store-bought
whey protein? If so, how many proteins can it resolve?
Independent Variable: 1 g, 2g, 3g of whey protein concentration
Dependent Variable: Number of bands shown in gel
Constant: Blank slot
Hypothesis:
We believe that within the whey protein are multiple amino acids because proteins are
made up of amino acids, and that they would show in different bands in the gel after
electrophoresis.
Introduction:
Our project is about using electrophoresis on whey protein to test and observe their
movement in an electrical current and also determining different size amino acids within the
protein by observing the bands that show on the gel.
Background information:
Electrophoresis is a lab technique that is used to separate proteins and nucleic acids in
size and charge then could be used for further examinations and observations of DNA. In
electrophoresis, molecules are placed in a gel and move towards its opposite charge, for example
a negatively charged molecule would move towards the positive end in the electric current while
a positively charged molecule would move negative side of the current. In addition, molecules
with a smaller mass would separate much quicker than molecules with a higher mass. The gels
that the molecules are placed in is called agarose gel. This gel is a polysaccharide extracted from
seaweed. Agarose gel is common in gel electrophoresis because they are very easy to prepare
and they are also non toxic. Within proteins are amino acids and the amino acids determine the
sequence in what the protein is functioned to do. Amino acids also determine the size, shape, and
charge of a protein. This project is interesting to us because within discoveries, there is always
electrophoresis involved. Electrophoresis is involved because determining its charge is size is
important in discovering what a protein is or what an DNA is. It may also be used in cancer
research because, looking at the result of a gel, comparing a cell to an cancerous cell, we can see
a difference in them by observing their lengths in how far they go in the gel, and how long it
takes them to move in the gel to determine what type of cell it is. The technique of gel
electrophoresis is now being used to solve many problems in evolution such as the amount and
type of genetic variation in populations and the genetic differences between species. The article
of University of California Press and American Institute of Biological Sciences describes the
merits of this new biochemical approach and examines some of the findings. We are interested in
this because we take what we like to drink which is protein shakes and using the process of
electrophoresis we were able to separate the supplement to see the different sizes of proteins and
if they are either negative or positively charged. Electrophoresis can be used during cancer
research to determine whether if a cell is cancerous by separating the proteins by size (cancer
cells being bigger than normal cells).
(http://www.jstor.org/discover/10.2307/1296112?uid=3739960&uid=2&uid=4&uid=3739256&s
id=21103315490997)
IV: Whey Protein
DV: Bands shown in gel
Constant: Blank slot
Methods:
Our hypothesis will be tested by putting protein into the gel box running at 200 volts for
50 minutes and by taking observations on the movement of the proteins and determining the
number of bands through a process of staining a gel after the 50 minutes. This process will begin
by getting a polyacrylamide gel and creating different compounds to be inserted into the gel. To
make a gel add 50 mL agarose, or other solutions such as polyacrylamide and the 950 mL buffer
solution and water and then heat it in the microwave for 60 seconds, interval of 2, 30 second runs.
After the gel is placed into the gel box, we pour in enough buffer solution that would make the
gel stay under the buffer. After the gel, we mix our compounds into different containers. In one
mix, we have 1 gram of whey protein mixed together with 10 mL of deionized water, another
with 2 gram of whey protein with 10 mL of deionized water, and 3 grams of deionized water. We
also have 1 gram, 2 grams, and 3 grams of dried milk with 10 mL of deionized water to add in
the empty slots in the gel. After the compounds is complete, we pipet in 10 mL of each solution
into each slot for its electrophoresis. The next step is to close the container and plug the positive
and the negative currents into opposite sides of the gel. The process of electrophoresis will take
about fifty minutes. After the gel is complete, we then start the staining process using Coomassie
blue stain in order to visualize the bands more accurately by having a container with the gel, and
leaving it in the Coomassie blue for one hour. After the staining process is the distaining process,
to remove the stain in order to represent the bands more accurately by leaving it with the distain
fluid for one extra hour. We record using pictures and qualitative data.
Procedure:

Prepare samples

Heat samples on heating block @ 70 degrees celsius for 10 min

Fill the gel upper buffer chamber with 200 mL running buffer, Fill lower chamber with
600 mL running buffer

Run Gel @ 200 Volts constant for 50 min

Observe and record
Materials:

Cylinder

Buffer solution

Water

Microwave

Whey Protein

Gel Box

10% PAGE (Gel)

50 mL 20x NuPAGE

950 mL DI water

Protein Standards

Coomassie blue stain
Results:
Gel:
3g Whey
2g Whey
1g Whey
3g Dried
2g Dried
1g Dried
Precision Blank
Protein
Protein
protein
Milk
Milk
Milk
Plus
concentration concentration concentration concentration concentration concentration Protein
Standard
Data: Electrophoresis
During the electrophoresis the compounds were moving at a fairly consistent rate. The
compounds with 1 grams of solute seemed to move a bit faster than the ones with 2grams and 3
grams. This is expected because during electrophoresis, smaller molecules would move faster in
the gel. After electrophoresis, little of the bands that were supposed to show were appearing.
This leads us to staining the gel using Coomassie blue to amplify the bands. After the staining
process was done, some compounds had distinct bands, while others had smeared bands.
Conclusion:
The data we got disproves our hypothesis, but it did answer part of our hypothesis. Although
bands were not visible to us, there are still proteins smeared onto the gel. Looking at our gel, we
can determine that proteins were definitely separated, but determining the amount is not possible
due to the smearing it caused. But working with protein electrophoresis, smearing is quite
common because proteins are bigger than DNA. This could be that, the amino acids that were
tested had to have multiple bands that were the same size, and multiple that were different sizes.
This could explain the reason why the bands are smeared, due to multiple factors in sizes, and
numbers of amino acids in whey protein.
Analysis:
Electrophoresis is used in the process of DNA separation by lengths. Using an electrical current
electrophoresis causes the DNA strands to move through the gel. Depending on how big and how
small the strands are allow them to move faster or slower through the gel. The smaller DNA
strands move faster through the gel than the bigger strands. Over time while the strands start to
work their way through the gel they start to separate by size allowing the experimenter to extract
certain groups of the DNA and examine each individually. This was important to research
because this will expand the use of electrophoresis, in this case it would expand it by identifying
the authenticity of weiy products. This experiment is an example of a person using our data in
the future by a person performing an experiment on the authenticity of a fast food restaurant’s
whey products. The only thing that would have improved the results is getting rid of any chance
of smearing by using something that contains less proteins of similar size.
References:
(2013)Principles of Gel Electrophoresis, Colostate.edu, Retrieved from
http://arbl.cvmbs.colostate.edu/
(2014) Retrieved from: Fred Hutch Cancer Research Center
N/A. (N/A). Gel electrophoresis. Retrieved from http://learn.genetics.utah.edu/content/labs/gel/
Schubert, F. (2013, August 16). Protein electrophoresis and multiple myeloma. Retrieved
from http://www.livestrong.com/article/337256-protein-electrophoresis-and-multiple-myeloma/
Rajkumar SV. Plasma cell disorders. In: Goldman L, Schafer AI, eds. Cecil Medicine. 24th ed.
Philadelphia, Pa: Saunders Elsevier; 2011:chap 193. Retrieved from: http://www.nlm.nih.gov/