We describe an undergraduate laboratory experiment in protein gel electrophoresis that uses readily available apparatus and materials. The separation of a mixture of stained proteins by gel electrophoresis was videotaped. Position–time data for the proteins generated from analysis of digitized videotape images allowed for calculation of protein terminal velocities. The dependence of protein terminal velocity on molar mass was determined and found to agree with predictions made by current theory. We also introduce a model that draws on simple physical concepts to help students place the experimental results in context.

1.
D.
Sheehan
,
Physical Biochemistry: Principles and Applications
(
Wiley
, Chichester,
2000
), Chap. 5;
C.
Viney
and
L. K.
Gilliland
, “
Gel electrophoresis of biological macromolecules
,” in
Modern Techniques for Polymer Characterization
, edited by
R. A.
Pethrick
and
J. V.
Dawkins
, (
Wiley
, Chichester,
1999
), Chap. 10.
Of background interest are
K.
Weber
,
J. R.
Pringle
, and
M.
Osborn
, “
Measurement of molecular weights by electrophoresis on SDS-acrylamide gel
,” in
Methods in Enzymology
, edited by
C. H. W.
Hirs
and
S. N.
Timasheff
(
Academic
, New York,
1972
),
Vol. XXVI-Enzyme Structure (Part C)
, pp.
3
27
;
A. L.
Shapiro
,
E.
Vinuela
, and
J. V.
Maizel
, Jr.
, “
Molecular weight estimation of polypeptide chains by electrophoresis in SDS-polyacrylamide gels
,”
Biochem. Biophys. Res. Commun.
28
,
815
820
(
1967
).
[PubMed]
2.
See, for example,
J. L.
Viovy
, “
Electrophoresis of DNA and other electrolytes: Physical mechanisms
,”
Rev. Mod. Phys.
72
,
813
872
(
2000
), and references therein.
3.
J.
Svasti
and
B.
Panijpan
, “
SDS-polyacrylamide gel electrophoresis
,”
J. Chem. Educ.
54
,
560
562
(
1977
);
K.
Weber
and
M.
Osborn
, “
Proteins and sodium dodecyl sulfate: Molecular weight determination on polyacrylamide gels and related procedures
,” in
The Proteins
, 3rd ed., edited by
H.
Neurath
and
R. L.
Hill
(
Academic
, New York,
1975
), Vol.
I
, Chap. 3;
J. A.
Reynolds
and
C.
Tanford
, “
The gross conformation of protein-sodium dodecyl sulfate complexes
,”
J. Biol. Chem.
245
,
5161
5165
(
1970
);
[PubMed]
K.
Weber
and
M.
Osborn
, “
The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis
,”
J. Biol. Chem.
244
,
4406
4412
(
1969
).
[PubMed]
4.
A.
Chrambach
and
D.
Rodbard
, “
Polyacrylamide gel electrophoresis
,”
Science
172
,
440
451
(
1971
).
5.
H. D.
Young
and
R. A.
Freedman
,
University Physics
, 10th ed. (
Addison–Wesley
, Reading,
2000
), Sec. 14-9.
6.
C.
Viney
and
R. A.
Fenton
, “
Physics and gel electrophoresis: Using terminal velocity to characterize molecular weight
,”
Eur. J. Phys.
19
,
575
580
(
1998
).
7.
Bio-Rad web site is http://www.bio-rad.com. Precast polyacrylamide gel: Tris-HCl Ready Gel (12% resolving gel, 4% stacking gel, 10 well, 30μl), product No. 161-1102. Vertical system chamber: Mini-PROTEAN II module, No. 165-2944. Buffer: Tris-HCl/glycine/SDS 10x concentrate, No. 161-0732. Protein mixture: Prestained precision protein standards, No. 161-0372 or No. 161-0373. Power supply: PowerPac 3000, No. 165-5056.
8.
S.
Erramilli
,
F.
Osterberg
, and
B.
Vogelaar
, “
Undergraduate laboratory: Principles of gel electrophoresis
,”
Am. J. Phys.
63
,
639
643
(
1995
).
9.
The public domain NIH Image program was developed at the U. S. National Institutes of Health and is available on the Internet at http://rsb.info.nih.gov/nih-image/.
10.
C.
Tanford
,
Physical Chemistry of Macromolecules
(
Wiley
, New York,
1961
), pp.
356
361
.
11.
P. G.
de Gennes
,
Scaling Concepts in Polymer Physics
(
Cornell University Press
, Ithaca,
1979
), Chap. VIII.
12.
S.
Bombard
,
A.
Favre
, and
J.
Kozelka
, “
Very short oligonucleotides migrate on 20% polyacrylamide gels with an apparent friction coefficient proportional to molecular mass
,”
Anal. Biochem.
263
,
123
125
(
1998
).
AAPT members receive access to the American Journal of Physics and The Physics Teacher as a member benefit. To learn more about this member benefit and becoming an AAPT member, visit the Joining AAPT page.