Figure 3.

Protein blot assays demonstrating specificity of ⁴⁵Ca²⁺ binding. Purified Dslo-C280 and various control proteins were subjected to SDS/PAGE, electroblotted onto poly(vinylidene difluoride) membrane, and processed for ⁴⁵Ca²⁺-overlay assay: (Right) image of exposed film; (Left) same blot stained with Coomassie Blue. (A) Lanes: 1, molecular mass markers; 2, Dslo-C280 (≈7 μg); 3, bovine calmodulin (4 μg); 4, rabbit calpain (6 μg); 5, FLAG-tagged E. coli alkaline phosphatase (4 μg); 6; bovine trypsin (4 μg); and 7, crude extract from E. coli expressing Dslo-C280. (B) Lanes: 1, Dslo-C280 (3 μg); 2, chicken troponin complex (6 μg); 3, human annexin V (3 μg); 4, Aspergillus niger glucose oxidase (3 μg); 5, glucose oxidase (6 μg); and 6, chicken ovomucoid protease inhibitor (3 μg).