Activator protein (AP), which stimulated fibroblast sphingomyelinase activity, was isolated from the spleen of a patient with Gaucher's disease type I by the combined techniques of heat and alcohol denaturation, DEAE-cellulose column chromatography, gel filtration, preparative polyacrylamide-gel electrophoresis and decyl-agarose chromatography. Urea/sodium dodecyl sulphate (SDS)/polyacrylamide-...

SDS is mostly used to enhance the solubilization and extraction of membrane proteins due to its strong detergency and low cost. Nevertheless, SDS interferes with the subsequent procedures and needs to be removed from the samples. In this work, a special gradient gel electrophoresis (GGE) system was developed to remove SDS from the SDS-solubilized protein samples. As a proof-of-principle experim...

We developed a method for the detection of phosphatase activity using fluorogenic substrates after polyacrylamide gel electrophoresis. When phosphatases such as Ca2+/calmodulin-dependent protein kinase phosphatase (CaMKP), protein phosphatase 2C (PP2C), protein phosphatase 5 (PP5), and alkaline phosphatase were resolved by polyacrylamide gel electrophoresis in the absence of SDS and the gel was...

1 The abbreviations used are: KDO, 3-Deoxy-D-manno-octulosonate; KDO 8-P, 3-DeoxyD-manno-octulosonate 8-phosphate; Tris, Tris-(hydroxymethyl)aminomethane; HEPES, N-[2Hydroxyethyl]piperazine-N’-[2-ethanesulfonic acid]; PNPase, purine nucleoside phosphorylase; EDTA, (ethylenedinitrilo)tetraacetic acid; 2D gel electrophoresis, two dimensional gel electrophoresis; SDS-PAGE, sodium dodecyl sulfate-p...

Analysis of proteins of the banana and citrus race of Radopholus similis was carried out by several different types of polyacrylamide gel electrophoresis. These included standard slab gel, SDS slab gel, gradient slab gel, and two-ditnensional slab gel electrophoresis. A major band difference was detected between the two races by slab gel electrophoresis. However, several other poorly resolved b...

SDS-polyacrylamide gel electrophoresis (SDS-PAGE) transfer method was used for purification and enrichment of the protein from crude sample. Coomassie bluc/ZnSO4 stained protein band(s) containing intact polyacrylamide gel were loaded on to another polyacrylamide gel either alone or as pooled gel bands. Two/three bands were combined together and arranged tightly over one another, sealed with st...

Volume 1 • Issue 7 • 1000e125 Biochem Anal Biochem ISSN:2161-1009 Biochem, an open access journal Electrophoresis is one of the research techniques, which has extensively been used in separation and identification of proteins/ enzymes. In majority of the biochemistry and molecular biology experiments, electrophoresis of proteins and enzymes is performed using polyacrylamide gel electrophoresis ...

Oxidative iodination of human lactoferrin (Lf) as commonly performed by using the chloramine-T, the Iodogen or the lactoperoxidase method produces an unreliable tracer protein because of excessive and heterogeneous polymer formation. Before iodination a minor tetramer fraction may be demonstrable in iron-saturated Lf only. Iodination-induced polymerization of iron-poor as well as iron-saturated...

The objective of this study was to characterize and compare the water and salt soluble proteins from rat liver and muscle by using sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and sodium dodecyl sulphate polymer-filled capillary gel electrophoresis (SDS-CGE). The evaluation and comparison of separations realized with the use of both techniques was done. It was found tha...