Tonin has been purified from rat submaxillary glands. The purification procedure included affinity chromatography on Sepharose 4B coupled to antitonin followed by DEAE chromatography and gel filtration on Sephadex G-100. Homogeneity of the purified enzyme was confirmed by Sephadex G-100 gel filtration, disc electrophoresis, and isoelectric focusing on polyacrylamide gel, immunodiffusion, and im...

Protein purification is essential for advancements in biotechnology. There are several different methods employed in purifying a particular protein from a complex sample such as a cell lysate. These methods take advantage of differences in the size, charge or binding affinity of the protein. One such method is affinity chromatography which utilizes the binding affinity of a protein toward a cer...

Many of the most widely employed operations in molecular biology hinge upon the use of single-stranded DNA as a probe or template. Here we report a straightforward method by which to produce long single-stranded DNA molecules using the polymerase chain reaction (PCR) in combination with immobilized metal affinity chromatography (IMAC). We demonstrate that a tag consisting of six successive 6-hi...

The purification of proteins by conventional procedures is frequently laborious and incomplete, and the yields are often low. Enzyme isolation based on a highly specific biological property-strong reversible association with specific substrates or inhibitors-has received only limited attention.‘-’ In affinity chromatography, the enzyme to be purified is passed through a column containing a cros...

A monocarboxamidomethyi derivative of human erythrocyte carbonic anhydrase B was purified by affinity chromatography. The modified enzyme possesses 3% of the COshydrating activity and 30% of the esterase activity of the native enzyme. The esterase activity is inhibited by the usual carbonic anhydrase inhibitors although the K; values are, in general, higher than for native enzyme. The pH depend...