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. 2015 Feb 4;15(1):16. doi: 10.1186/s12866-015-0352-5

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© Moscatiello et al.; licensee BioMed Central. 2015

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Isolation of acidic Ca ²⁺ buffering proteins from M . loti . Stains-all staining of SDS-PAGE (10-12.5% polyacrylamide linear gradient gel). Acidic proteins which bind Ca²⁺, such as calsequestrin and calreticulin, stain blue with Stains-all, whereas other proteins stain pink [21]. Lane 1, M. loti crude extracts (50 μg); lane 2, soluble fraction (50 μg); lane 3, protein sample after selective precipitation with ammonium sulphate (50 μg); lane 4, calsequestrin purified from rabbit skeletal muscle, used as a positive control (1 μg). Arrow, M. loti Stains-all blue-staining band with an apparent molecular mass of 20 kDa; arrowhead, rabbit calsequestrin.