Figure 5
From: A DNA aptamer population specifically detects Leishmania infantum H2A antigen
Specificity of the SELH2A aptamer population assessed by ELONA and Western blot assays. (a) Recombinant proteins H2A, H2B, H3, LIP0, LiP2a, LiP2b and BSA were plated at 2 μg/well (≈132 pmol/well) and incubated with 200 μl of 20 nM of digoxigenin-labeled SELH2A aptamer population. Afterwards, anti-digoxigenin-POD antibody was added and the plates were developed using ABTS solution. Abs 405 nm values were determined using a microplate reader from TECAN. All the experiments were made in triplicate and average of four different experiments is shown in the figure. Statistical significance with respect to the blank (a, P<0.001). Statistical significance between H2A and H3 (***P<0.001). (b) Thirty micrograms of total proteins (lane T) and nuclear proteins (lane N) from L. infantum (left panel or human cells (right panel) and 1 μg of recombinant H2A (lane R) were separated on SDS-15% PAGE gels and transferred to nitrocellulose membranes. Membranes were incubated with 1 μg/ml (40 nM) of the digoxigenin-labeled SELH2A aptamer population and probed with anti-digoxigenin-POD antibody. Finally, the membranes were developed with enhanced chemiluminescence's kits and exposed to hyperfilm. The sizes (in kDa) of the molecular size markers are indicated on the left. All the experiments were made in triplicate. Picture shows a representative experiment.
