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Cerium-based cytochemical method for detection of ouabain-sensitive, potassium-dependent p-nitrophenylphosphatase activity at physiological pH

T Kobayashi, T Okada and H Seguchi

We have developed a new cytochemical method for detecting the ouabain- sensitive, potassium-dependent p-nitrophenylphosphatase (K-NPPase) activity of the sodium-potassium-activated adenosine triphosphatase (Na- K ATPase) complex. The incubation medium contains p- nitrophenylphosphate (p-NPP) as substrate, cerium chloride as capture agent, Tricine buffer, MgCl2, and KCl. Tricine buffer protected against the medium turbidity caused by non-enzymatic reaction at pH 7.5. Biochemically, the accumulation of p-nitrophenol and phosphate in the reaction precipitate was proportionally related to the enzyme concentration. Ultracytochemically, the reaction products of the K- NPPase activity were localized as fine and uniform electron-dense deposits in the cytoplasmic side of specialized basolateral plasma membranes of cells of kidney distal convoluted tubules, secretory cells of salt gland, and marginal cells of stria vascularis. This method has the advantage of being useful at physiological pH.

Volume 35, Issue 5, pp. 601-611, 05/01/1987
Copyright © 1987 by The Histochemical Society


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Cytochemical Localization of Ouabain-sensitive, K+-dependent p-Nitrophenylphosphatase Activity in the Choroid Plexus of Normal and Reserpinized Guinea Pigs
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N. Kanoh
Cytochemical Localization of Ouabain-sensitive, K+-dependent p-nitrophenylphosphatase Activity in the Facial Nerve of Reserpinized Guinea Pigs
J. Histochem. Cytochem., August 1, 1997; 45(8): 1129 - 1136.
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