Universität zu Köln

Neumaier, F., Alpdogan, S., Hescheler, J. and Schneider, T. (2018). A practical guide to the preparation and use of metal ion-buffered systems for physiological research. Acta Physiol., 222 (3). HOBOKEN: WILEY. ISSN 1748-1716

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Abstract

Recent recognition that mobile pools of Zn2+ and Cu2+ are involved in the regulation of neuronal, endocrine and other cells has stimulated the development of tools to visualize and quantify the level of free trace metal ions. Most of the methods used to measure or control loosely bound metals require reference media that contain exactly defined free concentrations of the target ions. Despite the central importance of proper metal ion buffering, there is still a lack of international standards and beginners in the field may have difficulties finding a coherent description of how to prepare trace metal ion buffers, especially when experiments are to be performed in multimetal systems. To close this gap, we provide a guide for the design, preparation and use of metal ion-buffered systems that facilitate immediate application under physiologically relevant ionic conditions. Thermodynamic and kinetic concepts of chemical speciation as well as general protocols and specific examples are outlined for the accurate preparation of single- and dual-metal ion buffers. In addition, experiments have been performed with FluoZin-3 to illustrate that metal ion-buffered systems are required for reliable preparation of nanomolar Zn2+ solutions and that dual-metal ion buffers can be used to calibrate suitable fluorescent Zn2+ sensors in the presence of millimolar Ca2+ concentrations. Together, the information provided should sensitize readers to the many potential pitfalls and uncertainties that exist when working with physiologically relevant concentrations of trace metal ions and enable them to formulate their own metal ion buffers for most invitro applications.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Neumaier, F. UNSPECIFIED UNSPECIFIED UNSPECIFIED Alpdogan, S. UNSPECIFIED UNSPECIFIED UNSPECIFIED Hescheler, J. UNSPECIFIED UNSPECIFIED UNSPECIFIED Schneider, T. UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-195357
DOI:	10.1111/apha.12988
Journal or Publication Title:	Acta Physiol.
Volume:	222
Number:	3
Date:	2018
Publisher:	WILEY
Place of Publication:	HOBOKEN
ISSN:	1748-1716
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language APPARENT DISSOCIATION-CONSTANT; SUBSTITUTED AMINOSULFONIC ACIDS; CHEMICAL SPECIATION; LIGAND PURITY; GOODS BUFFERS; TRIS BUFFER; N-(2-ACETAMIDO)IMINODIACETIC ACID; COMPLEX FORMATION; PH BUFFERS; ZINC Multiple languages Physiology Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/19535