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Cerium-based cytochemical method for detection of ouabain-sensitive, potassium-dependent p-nitrophenylphosphatase activity at physiological pH.
J Histochem Cytochem. 1987 May; 35(5):601-11.JH

Abstract

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.

Authors

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Pub Type(s)

Journal Article

Language

eng

PubMed ID

3031156

Citation

Kobayashi, T, et al. "Cerium-based Cytochemical Method for Detection of Ouabain-sensitive, Potassium-dependent P-nitrophenylphosphatase Activity at Physiological PH." The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society, vol. 35, no. 5, 1987, pp. 601-11.
Kobayashi T, Okada T, Seguchi H. Cerium-based cytochemical method for detection of ouabain-sensitive, potassium-dependent p-nitrophenylphosphatase activity at physiological pH. J Histochem Cytochem. 1987;35(5):601-11.
Kobayashi, T., Okada, T., & Seguchi, H. (1987). Cerium-based cytochemical method for detection of ouabain-sensitive, potassium-dependent p-nitrophenylphosphatase activity at physiological pH. The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society, 35(5), 601-11.
Kobayashi T, Okada T, Seguchi H. Cerium-based Cytochemical Method for Detection of Ouabain-sensitive, Potassium-dependent P-nitrophenylphosphatase Activity at Physiological PH. J Histochem Cytochem. 1987;35(5):601-11. PubMed PMID: 3031156.
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TY - JOUR T1 - Cerium-based cytochemical method for detection of ouabain-sensitive, potassium-dependent p-nitrophenylphosphatase activity at physiological pH. AU - Kobayashi,T, AU - Okada,T, AU - Seguchi,H, PY - 1987/5/1/pubmed PY - 1987/5/1/medline PY - 1987/5/1/entrez SP - 601 EP - 11 JF - The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society JO - J Histochem Cytochem VL - 35 IS - 5 N2 - 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. SN - 0022-1554 UR - https://www.unboundmedicine.com/medline/citation/3031156/Cerium_based_cytochemical_method_for_detection_of_ouabain_sensitive_potassium_dependent_p_nitrophenylphosphatase_activity_at_physiological_pH_ L2 - https://journals.sagepub.com/doi/10.1177/35.5.3031156?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub=pubmed DB - PRIME DP - Unbound Medicine ER -