Abstract
The method of purification of Na,K-ATPase from pig kidney is based on a differential centrifugation and SDS-treatment of a microsomal preparation. The yield is 0.4 mg protein per 1 g tissue with the specific (ouabain-sensitive) activity of 25–28 μmol Pi/min per mg protein and nucleotide binding capacity of 3 nmol/mg. The protein/lipid ratio is 1/1 (mg/mg) with a protein purity of ~80 %.
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References
Methods in Enzymology 156, Section I. Preparation of Na+,K+-ATPase and Subunits. pp. 29–71
Klodos I, Esmann M, Post RL (2002) Large scale preparation of sodium-potassium ATPase from kidney outer medulla. Kidney Int 62:2097–2100
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Acknowledgement
I thank Dr. Mikael Esmann for helpful suggestions. Ms. Birthe Bjerring Jensen, Anne Lillevang, and Angelina Damgaard are gratefully acknowledged for their expert advice and practical tips.
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Fedosova, N.U. (2016). Purification of Na,K-ATPase from Pig Kidney. In: Bublitz, M. (eds) P-Type ATPases. Methods in Molecular Biology, vol 1377. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3179-8_2
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DOI: https://doi.org/10.1007/978-1-4939-3179-8_2
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3178-1
Online ISBN: 978-1-4939-3179-8
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