It has been demonstrated by several investigators, most convincingly by studies of the effect of urea on the solubility of various compounds, that aqueous urea solutions are better solvents than pure water for many solutes containing hydrophobic groups (l-5). It has further been suggested that this property of aqueous urea solutions is responsible for their denaturing activity toward proteins a...

Denaturing Isoelectric Focusing Free-Flow Electrophoresis (IEF-FFE) Fractionation of Crude Protein Mixtures Peter J.A. Weber, Gerhard Weber, and Christoph Eckerskorn This protocol was adapted from "Denaturing IEF-FFE Fractionation of Crude Protein Mixtures," contributed by Peter J.A. Weber, Gerhard Weber, and Christoph Eckerskorn, Chapter 19, in Purifying Proteins for Proteomics (ed. Simpson). ...

60-70S RNA of avian myeloblastosis virus was spread with the Kleinschmidt method and examined in the electron microscope after being subjected to a variety of denaturing conditions. Extension of the molecules from a collapsed configuration was seen only under stringent denaturing conditions, resulting in molecules of 1-2 mum in length. Longer strands of these extremely fragile molecules cannot ...

Single-strand conformational polymorphism and denaturing gel electrophoresis were used to screen for mutations in the protoporphyrinogen oxidase gene (PPOX) of three patients with clinically and biochemically proven variegate porphyria in order to select genomic regions for specific DNA sequence analysis. Two previously undescribed mutations were identified: PPOX1423-1426-delATCT and PPOX2272in...

Western blotting is an analytical technique used to detect specific proteins in a given sample of tissue homogenate or extract. It uses gel electrophoresis to separate native or denatured proteins by the length of the polypeptide (denaturing conditions) or by the 3-D structure of the protein (native/ non-denaturing conditions). The proteins are then transferred to a membrane (typically nitrocel...

We used denaturing gradient gel electrophoresis to detect the beta-thalassemia mutations in the Chinese population. By amplifying the beta-globin gene in four separate fragments and electrophoresing the amplified DNA in two gels, we were able to distinguish all the 12 known mutations on the basis of the mobility of the homoduplexes and heteroduplexes. We conclude that denaturing gradient gel el...