Gel electrophoresis

About: Gel electrophoresis is a research topic. Over the lifetime, 26026 publications have been published within this topic receiving 1113565 citations.

Highly Sensitive and Fast Protein Detection with Coomassie Brilliant Blue in Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis

Abstract: Department of Chemistry, Sungkyunkwan University, Suwon 440-746, KoreaReceived August 12, 2002Key Words : Coomasie brilliant blue, 2-Dimensional gel electrophoresis, ProteomicsDetection of proteins in sodium dodecyl sulfate-polyacryl-amide gel electrophoresis (SDS-PAGE) is an important firststep for protein analysis. For years, various experimentalefforts have been directed to develop and improve theprotein detection methods

Identification and purification of glial growth factor

Abstract: Cultured rat Schwann cells are stimulated to divide by a protein growth factor, present in extracts of bovine brain and pituitary, which we have named glial growth factor (GGF). Two lines of evidence indicate that GGF activity in both brain and pituitary resides in a protein of Mr = 31,000. (1) Four independently isolated monoclonal antibodies that immunoprecipitate the activity react with an antigen of this molecular weight in sodium dodecyl sulfate (SDS)-polyacrylamide gels. (2) After SDS-polyacrylamide gel electrophoresis of partially purified preparations, mitogenic activity on Schwann cells is recovered at this molecular weight. GGF has been purified approximately 10(5)-fold to apparent homogeneity from bovine pituitary anterior lobes by a combination of column chromatography steps and preparative SDS gel electrophoresis. Purified human platelet-derived growth factor, a molecule with properties similar to those of GGF, is inactive on Schwann cells and therefore appears to be distinct.

A brain protein unique to the olfactory bulb.

Abstract: Soluble extracts from several regions of the mouse brain manifested regionally specific protein band patterns on polyacrylamide gel electrophoresis. In particular, one protein band appeared to occur uniquely in the olfactory bulb extracts where it was a quantitatively significant constituent of the soluble protein extract. This protein was purified to homogeneity by ammonium sulfate precipitation, DEAE-cellulose column chromatography, and polyacrylamide gel electrophoresis. The purified protein has a minimum molecular weight of about 20,000 by gel electrophoresis in the presence of sodium dodecyl sulfate. Precipitating antiserum prepared against this protein reacted only with the purified protein or with extracts of olfactory bulb, and not with extracts of other mouse tissues or brain regions. Quantitative immunoprecipitation assays indicate that this specific protein represents 1% of the total soluble protein in the mouse olfactory bulb. Soluble extracts of olfactory bulbs from several rodent species gave reactions of identity by agar gel diffusion with the antiserum to the mouse protein. It is suggested that this protein is an example of selective genetic expression within the central nervous system.

Immunochemical identification of membrane proteins after sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

Abstract: Publisher Summary Electrophoresis in the presence of sodium dodecyl sulfate (SDS) is a key method for analyzing membrane proteins, and provides information on the size and the amount of these molecules. This chapter describes a method, immunochemical identification of membrane proteins after sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and discusses its various possible applications in the study of membrane proteins. These procedures are based on the transfer of proteins from the polyacrylamide gel slab to an insoluble matrix, such as chemically reactive cellulose or, more conveniently, commercially available nitrocellulose sheets. The immobilized proteins on the replica are readily accessible to antibodies or other specific probes; this makes possible the in situ localization and characterization of individual proteins in complex mixtures. Polypeptides transferred to nitrocellulose sheets can also be identified with specific probes other than antibodies—for example, glycoproteins can be visualized by reaction with radio-labeled lectins, such as concanavalin A or wheat germ agglutinin.