THE JOURNAL OF BIOLOGICAL CHEMISTRY Vol. 281, No. 24, Issue of June 16, p.

e19, 2006
© 2006 by The American Society for Biochemistry and Molecular Biology, Inc. Printed in U.S.A.

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A PAPER IN A SERIES REPRINTED TO CELEBRATE THE CENTENARY OF THE JBC IN 2005

JBC Centennial
1905–2005
100 Years of Biochemistry and Molecular Biology

SDS-PAGE to Determine the Molecular Weight of Proteins:

the Work of Klaus Weber and Mary Osborn
The Reliability of Molecular Weight Determinations by Dodecyl Sulfate-
Polyacrylamide Gel Electrophoresis
(Weber, K., and Osborn, M. (1969) J. Biol. Chem. 244, 4406 – 4412)
This Journal of Biological Chemistry (JBC) Classic on using sodium dodecyl sulfate-poly-
acrylamide gel electrophoresis (SDS-PAGE) to determine the molecular weight of proteins is
one of our most highly cited articles. According to the Thomson Scientific Web of Science it was
the 13th most cited article in 2004, with 23,167 total citations. It was also the fourth most cited
paper between 1945 and 1988, with 20,672 citations. The paper comes from the husband and
wife team of Klaus Weber and Mary Osborn who were both at Harvard University at the time
the paper was published.
Weber was an Assistant Professor in Harvard’s Department of Biochemistry and Molec-
ular Biology. He was originally from Poland, born in Lodz in 1936. After earning his
undergraduate (1962) and graduate (1964) degrees from the University of Freiburg, Weber
came to the United States to work with James Watson at Harvard. He was eventually hired
as an Assistant Professor and ran a joint laboratory with Watson and Walter Gilbert.
Osborn, who was born in Darlington, England in 1940 and attended Cambridge University
(B.A. in 1962) and Pennsylvania State University (Ph.D. in 1967), was a research fellow in
the laboratory.
When the paper was published in 1969, the molecular weights of proteins were determined
by a variety of techniques, all of which were quite experimentally demanding. Two years
earlier, Shapiro, Vinuela, and Maizel (1) reported that proteins could be separated by poly-
acrylamide electrophoresis in the presence of sodium dodecyl sulfate and that this separation
was dependent on the molecular weights of the polypeptide chains. In 1968, Weber used this
method to determine the molecular weights of aspartate transcarbamylase from Escherichia
coli (2). His successful use of this technique made Weber wonder how widely applicable the
method was.
To test the utility of electrophoresis for determining the molecular weights of proteins,
Weber and Osborn selected 40 proteins with known molecular weights. They ran the proteins
on gels and plotted their electrophoretic mobilities against the logarithm of their molecular
weights. They found that a smooth curve was produced, even when they used proteins that
were globular and proteins that were highly helical, rod-shaped molecules. Weber and Osborn
concluded that “the good resolution and the fact that an estimate of the molecular weight can
be obtained within a day, together with the small amount of protein needed, makes the method
strongly competitive with others commonly employed.” They suggested that the technique
could be used with commercially available proteins as standards to determine the molecular
weights of polypeptide chains.
Weber eventually became a Full Professor at Harvard (1972), and Osborn took a staff
position first at the Laboratory of Molecular Biology in Cambridge (1969 –1972) and then at
Cold Spring Harbor Laboratory (1972–1975). The pair eventually moved to Germany in 1975
when Weber was offered the position of Director of the Department of Biochemistry and Cell
Biology at the Max Planck Institute for Biophysical Chemistry in Göttingen. There they
This paper is available on line at http://www.jbc.org e19

This is an Open Access article under the CC BY license.

e20 Classics

Klaus Weber and Mary Osborn. Photo courtesy of Klaus Weber.

pioneered another new technique: immunofluorescence microscopy. They found that they
could tag microtubules with antibodies and then tag these antibodies with a second fluorescent
antibody. The light emitted by the antibodies made the microtubules visible throughout the
cell. This technique could also be modified to produce color pictures of whole cells.
Currently, Osborn and Weber remain at the Max Planck Institute where they run a joint
laboratory that focuses on the cytoskeletal elements of mammalian cells and networks gov-
erning cellular architecture. Weber is Emeritus Director of the Institute and Professor at the
University of Göttingen. Osborn is an Honorary Professor at the University of Göttingen and
also the current President of the International Union of Biochemistry and Molecular Biology
(IUBMB).
In recognition of their work, Weber and Osborn have received many honors and awards.
Weber was awarded the Ernst Jung Prize for Medicine in 1984, the Otto-Warburg Medal from
the German Society for Biochemistry and Molecular Biology in 1997, and the Carl Zeiss Prize
from the German Society of Cell Biology, which he shared with Osborn in 1998. Weber has also
served on the Editorial Boards of Cell, EMBO Journal, Experimental Cell Research, the
European Journal of Cell Biology, and Mechanisms of Development. Osborn was awarded the
Meyenburg Prize for Cancer Research in 1987, the Helena Rubenstein/UNESCO Prize for
Women in Science in 1998, and the L’Oreal/UNESCO Prize for Women in Science in 2002. She
has served on the Editorial Boards of Cell, Experimental Cell Research, Differentiation,
Seminars in Cancer Biology, Biology of the Cell, and Subcellular Biochemistry.
Nicole Kresge, Robert D. Simoni, and Robert L. Hill
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REFERENCES
1. Shapiro, A. L., Vinuela, E., and Maizel, J. V., Jr. (1967) Molecular weight estimation of polypeptide chains by
electrophoresis in SDS-polyacrylamide gels. Biochem. Biophys. Res. Commun. 28, 815– 820
2. Weber, K. (1968) New structural model of E. coli aspartate transcarbamylase and the amino-acid sequence of the
regulatory polypeptide chain. Nature 218, 1116 –1119