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.
Hoepflinger et al. BMC Plant Biology 2013 13:135 doi:10.1186/1471-2229-13-135