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...

We describe a polyacrylamide gel casting cassette that overcomes limitations of commercially available gel electrophoresis equipment. This apparatus molds a single polyacrylamide gel that can evaluate more than 200 samples in parallel, is loaded with a multichannel pipettor, and is flexible with respect to composition of the separating matrix. We demonstrate its use to characterize inhibitors o...

We demonstrate a two-dimensional microfluidic architecture that integrates polyacrylamide gel electrophoresis (PAGE) with immunoblotting in a fully automated format. This assay is designed to overcome performance limitations of conventional slab-gel immunoblotting, including multiple manual interventions, low-throughput transfer and blotting, and substantial consumption of reagents and sample. ...

Four of the membrane proteins from Acholeplasma laidlawii that are soluble in the nonionic detergent Tween 20 have been purified by preparative electrophoretic techniques utilizing different supporting media. The last purification step for two of the major proteins was a preparative polyacrylamide gel electrophoresis performed in the absence of any detergent. The proteins were recovered by cont...

A thermostable alkaline protease was isolated from the Aspergillus niger Z1 strain in a liquid Czapek Dox medium, containing casein (1% w/v) as the sole nitrogen source. Enzyme extract was subjected to electrophoresis in SDS-polyacrylamide gel. Two protein bands were seen on polyacrylamide gel. Active enzyme was visualized in a zymogram and protease activity exhibited a molecular mass of 68 kDa...

Renin was purified from 47 kg of hog kidney to produce enough enzyme for enzymatic and physicochemical characterization. The procedure included extraction at pH 3.5 in the presence of protease inhibitors, two ammonium sulfate precipitations, ion exchange chromatography on Sepharose-hexamethylenediamino-pepstatin gel, gel filtration, and isoelectric focusing. Renin, 2.3 mg, with a specific activ...