| Assay Method Information | |
| | Test for Activity of Compounds in Inhibiting Lp(a) Assembly |
| Description: | Experimental steps: Equal amounts of Apo(a) and ApoB serum were mixed with a serially diluted test compound (highest concentration at 100 nM, serially diluted 3-fold). The mixture was incubated in a 37° C. incubator for 2 h. The reaction was then terminated by adding 6-aminocaproic acid (EACA, purchased from Sigma) to a final concentration of 150 mM. The post-reaction solution was then added to an ELISA plate pre-coated with ApoB-Capture antibody, and the plate was incubated at room temperature for 2 h. Then the plate was washed 4 times with wash buffer; biotinylated Apo(a)-Detector antibody was added, and the plate was incubated at room temperature for 1 h. The plate was washed, and the chromogenic substrate 3,3′,5,5′-tetramethylbenzidine (TMB, purchased from Abcam) solution was added. After the plate was incubated at room temperature for 15 min, a reaction stop solution was added. Immediately after the solution was mixed well, the absorbance at 450 nm was measured using a microplate reader. Finally, data analysis and IC50 calculation were performed using GraphPad Prism 9 software. |
| Affinity data for this assay | |
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