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-...

development of a stool antigen immunoassay to detect helicobacter pylori infection requires monoclonal antibody against the specific antigen. alkylhydroperoxide reductase (ahpc) of helicobacter pylori has been described as a specific and unique enzyme for h. pylori and therefore, both h. pylori ahpc and anti-ahpc could be useful in the development of serologic and stool antigen tests, for detec...

1Plant Molecular and Cellular Genetics Lab, School of Agriculture and Home Economics, Tuskegee University, Tuskegee, Alabama 36088-1614; 2USDA/ARS, Plant Introduction Station, Griffin, Georgia 30223; 3Department of Plant Pathology, Auburn University, Alabama 36849 Agarose is the preferred matrix for resolving DNA fragments resulting from PCR amplifications using arbitrary primers for the detect...

Designed for compatibility with slab-gel polyacrylamide gel electrophoresis (PAGE) reagents and instruments, we detail development of free-standing polyacrylamide gel (fsPAG) microstructures supporting electrophoretic performance rivalling that of microfluidic platforms. For the protein electrophoresis study described here, fsPAGE lanes are comprised of a sample reservoir and contiguous separat...

A simple method for preparation and fractionation of the streptococcal nucleases by polyacrylamide gel electrophoresis is presented. The procedure is carried out with ammonium sulfate-precipitated supernatant fluids from cultures of beta-hemolytic streptococci grown to stationary phase. Electrophoresis on polyacrylamide gel and subsequent elution of the fractionated enzymes allows the preparati...

We describe a procedure for the convenient separation of proteins by sodium dodecyl sulfate/polyacrylamide gel electrophoresis of urine from cases of renal disease. A precipitation method that requires no special apparatus was used to concentrate the urinary proteins; for electrophoretic separation we used a commercially supplied polyacrylamide/cellulose gel slab. This method seems to be valuab...

b-cyclodextrin glucosyl transferase (b-cgtase) hydrolyses starch to produce b-cyclodextrin by transglycosylation (cyclization) activity. the conventional method for detection of b-cgtase activity is based on detecting starch hydrolysis by iodine staining. this method reveals all amylolytic enzymes, but can not discriminate them. in the present study, we introduce a new method for specific detec...

A hybridoma cell line was isolated which produced monoclonal antibody to one protein component of a yeast-phase cytoplasmic antigenic complex of Blastomyces dermatitidis. The immunoglobulin M antibody product was characterized by immunodiffusion, autoradiography of polyacrylamide gels, and cellulose acetate electrophoresis. By attaching the antibody to an affinity gel, one major protein band wa...

We have demonstrated that the gel shift assay, a powerful method to study protein.DNA interactions under equilibrium conditions, is both an accurate and precise method to measure the affinity of anti-DNA.DNA immune complexes. One difficulty in performing gel shift assays is disruption of protein.DNA equilibria during the time needed for complexes to enter the gel matrix. However, we have found ...

Uroporphyrinogen III synthase purified from rat liver is a monomer of Mr 36,000 by gel filtration and 28,000 by SDS/polyacrylamide-gel electrophoresis. The enzyme exists in two interconvertible forms separable on h.p.l.c. Both forms of the enzyme could be renatured with full activity after SDS/polyacrylamide-gel electrophoresis, demonstrating the absence of a reversibly bound cofactor. The enzy...