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. 2013 Sep 16;13:135. doi: 10.1186/1471-2229-13-135

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Copyright © 2013 Hoepflinger et al.; licensee BioMed Central Ltd.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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⁴⁵Ca overlay assay of recombinant AtTCTP. Ca²⁺-binding assay using radiolabelled calcium and recombinant AtTCTP. A shows a western blot of recombinant StrepII-tagged AtTCTP (MW = protein marker, AtTCTP = recombinant AtTCTP-StrepII; important marker band sizes are indicated). B shows the ⁴⁵Ca overlay assay of AtTCTP, aequorin (aeq), RuBisCo (RBC), bovine serum albumin (BSA), and controls C1 (10 μl purified crude extract without StrepII-tag protein expression) and C2 (non Ca²⁺-binding, StrepII-tagged A. thaliana protein kinase). C shows Coomassie brilliant blue staining of the PVDF membrane used in 3B. If not indicated differently, 0.25 μg of each protein were spotted for ⁴⁵Ca overlay assay.