One-dimensional SDS gel electrophoresis of proteins

Current Protocols in Immunology
Sean R Gallagher

Abstract

Electrophoresis is used to separate complex mixtures of proteins (e.g., from cells, subcellular fractions, column fractions, or immunoprecipitates), to investigate subunit compositions, and to verify homogeneity of protein samples. It can also serve to purify proteins for use in further applications. In polyacrylamide gel electrophoresis, proteins migrate in response to an electrical field through pores in a polyacrylamide gel matrix; pore size decreases with increasing acrylamide concentration. The combination of pore size and protein charge, size, and shape determines the migration rate of the protein. In this unit, the standard Laemmli method is described for discontinuous gel electrophoresis under denaturing conditions, that is, in the presence of sodium dodecyl sulfate (SDS). Both full-size and minigel formats are detailed. Several modifications are provided for specific applications. For separation of peptides and small proteins, the standard buffers are replaced with either a Tris-tricine buffer system or a modified Tris buffer in the absence of urea. Continuous SDS-PAGE is a simplified method in which the same buffer is used for both the gel and electrode solutions and the stacking gel is omitted. Other protocols cover ...Continue Reading

References

Jul 1, 1978·Analytical Biochemistry·P T Matsudaira, D R Burgess
Jan 1, 1972·Methods in Enzymology·K WeberM Osborn
Mar 27, 2007·Cellular Microbiology·Daniel E Voth, Robert A Heinzen
Apr 23, 2008·Current Protocols in Protein Science·H L Ploegh

❮ Previous
Next ❯

Citations

Feb 12, 2008·Current Protocols in Molecular Biology·Carmen A Argmann, Johan Auwerx
Oct 6, 2011·Current Protocols in Neuroscience·Markus U EhrengruberKenneth Lundstrom
Aug 2, 2016·Current Protocols in Immunology·Duojiao NiSean Gallagher
Jan 30, 2008·Current Protocols in Cell Biology·Kevin L LorickAllan M Weissman
Apr 4, 2017·Current Protocols in Immunology·Amrendra KumarSebastian Joyce
Nov 2, 2016·Current Protocols in Immunology·John D Altman, Mark M Davis
Sep 5, 2019·Current Protocols in Immunology·Jolien J LuimstraHuib Ovaa
Aug 24, 2011·Journal of Agricultural and Food Chemistry·Mikhail F BorisenkovFedor V Vityazev
Dec 7, 2021·Current Protocols·Collin LaakerWilliam J Karpus
Dec 11, 2021·Current Protocols·Edward F RosloniecDavid D Brand

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.

Related Papers

Current Protocols in Cell Biology
Sean R Gallagher
Current Protocols in Toxicology
Sean R Gallagher
Current Protocols in Molecular Biology
Sean R Gallagher
Journal of Visualized Experiments : JoVE
Bulbul Chakavarti, Deb Chakavarti
Current Protocols in Protein Science
Sean R Gallagher
© 2022 Meta ULC. All rights reserved