This account gives an overview and evaluation of the current methods of most commonly used electrophoretic techniques in the analysis of various polypeptides. A general review of some background information and pitfalls associated with the application of these methods in the characterization of the size and charge properties of proteins or peptides is presented. It is intended to provide some g...

Many protozoan parasites represent an important group of human pathogens. Pulsed Field Gradient Gel Electrophoresis (PFGE) analysis has been an important tool for fundamental genetic studies of parasites like Trypanosoma, Leishmania, Giardia or the human malaria parasite Plasmodium falciparum. We present PFGE conditions allowing a high resolution separation of chromosomes ranging from 500 to 40...

Two-dimensional gel electrophoresis has been instrumental in the development of proteomics. Although it is no longer the exclusive scheme used for proteomics, its unique features make it a still highly valuable tool, especially when multiple quantitative comparisons of samples must be made, and even for large samples series. However, quantitative proteomics using 2D gels is critically dependent...

In this study, we examined yeast proteins by two-dimensional (2D) gel electrophoresis and gathered quantitative information from about 1,400 spots. We found that there is an enormous range of protein abundance and, for identified spots, a good correlation between protein abundance, mRNA abundance, and codon bias. For each molecule of well-translated mRNA, there were about 4,000 molecules of pro...

The two-dimensional blue native/sodium dodecyl sulfate polyacrylamide gel electrophoresis (2D BN/SDS-PAGE) is a method of choice for the investigation of protein complexes. This highly resolvent separation method is unique in that it facilitates the identification of many protein complexes simultaneously. Because of its simplicity and suitability, the 2D BN/SDS-PAGE can be now applied to a wide...

We utilized two-dimensional gel electrophoresis and immunoblotting (2D-immunoblotting) with anti-Sporothrix schenckii antibodies to identify antigenic proteins in cell wall preparations obtained from the mycelial and yeast-like morphologies of the fungus. Results showed that a 70-kDa glycoprotein (Gp70) was the major antigen detected in the cell wall of both morphologies and that a 60-kDa glyco...

We have modified one of the most useful methods of protein separation; namely, two dimensional gel electrophoresis (2-DE). This modified version of 2-DE is not only simpler and easier but also faster than all the currently available methods. In this method, isoelectric focusing is carried out in the first dimension using a vertical sodium dodecyl sulfate polyacrylamide gel electrop...

MATERIALS 6x Gel-loading buffer Agarose solutions (please see Step 3) DNA samples DNA size standards Samples of DNAs of known size are typically generated by restriction enzyme digestion of a plasmid or bacteriophage DNA of known sequence. Alternatively, they are produced by ligating a monomer DNA fragment of known size into a ladder of polymeric forms. DNA staining solution For a discussion of...

Gel electrophoresis is a widely known group of techniques used to separate and identify macromolecules as DNA, RNA, or proteins based on size, form, or isoelectric point. The separation of molecules by electrophoresis is based on the fact that charged molecules migrate through a gel matrix upon application of an electric field. These techniques have become a main tool in biochemistry, molecular...

Agarose gel electrophoresis is used to separate DNA or RNA molecules based upon their size.