Turning a PAGE: the overnight sensation of SDS-polycrylamide gel electrophoresis

Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, 377 Plantation St., Worcester, MA 01605, USA.
The FASEB Journal (Impact Factor: 5.48). 05/2008; 22(4):949-53. DOI: 10.1096/fj.08-0402ufm
Source: PubMed

ABSTRACT The zonal separation of proteins on the basis of net charge was initially conducted on paper, then in columns of sucrose and later in gels of starch and polyacrylamide, with appropriate electric fields. Then, in 1964, a graduate student at MIT discovered the power of sodium dodecyl sulfate (SDS) to dissociate the envelope proteins of Escherichia coli and to dramatically enhance their electrophoretic resolution when the detergent was included in the gel. While this Ph.D. thesis work continued, a group at the Albert Einstein College of Medicine published in 1965 the use of SDS to disrupt poliovirus particles and to resolve the proteins in gels containing SDS. This group soon followed with a publication (1966) on the application of this new method to the study of immunoglobulin heavy and light chain synthesis. Because of concurrent advances in gel filtration and other methods of protein separation, SDS gel electrophoresis had its greatest impact not in biochemistry but in cell biology and virology. Ingenious devices were soon introduced that facilitated the application of this method to radioactive protein mixtures, followed by the introduction of slab gels for the simultaneous resolution of multiple samples in parallel lanes in a single run. As we today routinely perform "SDS PAGE" (as the method become known, to the great irritation of journal copyeditors and nomenclature committees at the time), it is fitting to pause--four decades later, and remember the pioneers who made SDS gel electrophoresis a reality, a true milestone that caught on almost overnight.

  • Source
    Encyclopedia of Analytical Chemistry, 09/2009; , ISBN: 9780470027318
  • [Show abstract] [Hide abstract]
    ABSTRACT: The nucleolus is among the most striking cell components when viewed by phase contrast microscopy and yet, recent studies have revealed it to possess a lower molecular density than anticipated. A new report demonstrates that the nucleolus accommodates the assembly of a virus, this amidst building ribosomes and performing other host cell functions.
    Nucleus (Austin, Texas) 01/2010; 1(5):444-5. DOI:10.4161/nucl.1.5.13056 · 3.15 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: SDS/PAGE is universally used in biochemistry, cell biology, and immunology to resolve minute protein amounts readily from tissue and cell extracts. Although molecular weights of water-soluble proteins are reliably determined from their SDS/PAGE mobility, most helical membrane proteins, which comprise 20-30% of the human genome and the majority of drug targets, migrate to positions that have for decades been unpredictably slower or faster than their actual formula weight, often confounding their identification. Using de novo designed transmembrane-mimetic polypeptides that match the composition of helical membrane-spanning sequences, we quantitate anomalous SDS/PAGE fractionation of helical membrane proteins by comparing the relative mobilities of these polypeptides with typical water-soluble reference proteins on Laemmli gels. We find that both the net charge and effective molecular size of the migrating particles of transmembrane-mimetic species exceed those of the corresponding reference proteins and that gel acrylamide concentration dictates the impact of these two factors on the direction and magnitude of anomalous migration. Algorithms we derived from these data compensate for this differential effect of acrylamide concentration on the SDS/PAGE mobility of a variety of natural membrane proteins. Our results provide a unique means to predict anomalous migration of membrane proteins, thereby facilitating straightforward determination of their molecular weights via SDS/PAGE.
    Proceedings of the National Academy of Sciences 09/2013; DOI:10.1073/pnas.1311305110 · 9.81 Impact Factor


1 Download
Available from