A Histochemical Method for the Demonstration of Diphosphopyridine Nucleotide Diaphorase

A Histochemical Method for the Demonstration of Diphosphopyridine Nucleotide Diaphorase

The present investigation concerning the histochemical demonstration of DPN diaphorase follows the development of a new reagent, Nitro-BT, which has already been used successfully for the cytochemical localization of the succinic dehydrogenase system. The most consistently favorable results were obtained with the lactate-lactic dehydrogenase system buffered at pH 7.4. Using sections of rat kidn...

The Histochemical Localization of Triphosphopyridine Nucleotide Diaphorase

A histochemical method is described for the localization of triphosphopyridine nucleotide diaphorase using a recently synthesized tetrazolium salt (Nitro-BT). By virtue of the favorable histochemical properties of this reagent, it has been possible to demonstrate that whereas DPN diaphorase is usually restricted to the mitochondria, the TPN diaphorase activity of corresponding cells was distrib...

Purification and properties of diphosphopyridine nucleotide diaphorase of human erythrocytes.

Diaphorase Assay-Six-hundredths pmole of 2,6-dichlorophenolindophenol, 50 pmoles of Tris-HCl, pH 7.55, 1 pmole of EDTA, enzyme, and 0.44 pmole of DPNH in a volume of 3 ml were placed in a Beckman spectrophotometer equipped with a water jacket through which water at 25” was circulated. A control cuvette without enzyme was included, and absorbancy at 600 rnp was determined for 10 minutes. Under t...

A Micro Manometric Method for the Determination of Diphosphopyridine Nucleotide

In the course of histochemical studies of the retina, it was desired to determine the diphosphopyridine nucleotide content of single microtome sections of tissue weighing on the order of 10 to 50 y. It was, therefore, necessary to increase the sensitivity of the manometric method for the determination of DPN. To this end, the technique of Linderstrom-Lang (6) with the Cartesian diver was employ...

A Manometric Method for the Determination of Diphosphopyridine Nucleotide