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Assay Method Information |
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| | PCSK9 Inhibition Assay |
| Description: | For the peptide affinity determinations, biotinylated PCSK9 was immobilized to NeutrAvidin-coated MaxiSorp Immuno-Plates and incubated with a mixture of serially diluted synthetic peptide (Pep2-2 or Pep2 at 0-100 μM) and the correspondingpeptide-displaying phage at the earlier determined subsaturating concentration (see above). After a 1-h incubation in PBT buffer (PBS, 0.5% BSA, 0.05% Tween 20), the plate was washed extensively, and the bound phage was detected by anti-M13-HRP. The data were fitted to a four-parameter equation (KaleidaGraph) to determine the IC50 values. For IC50 measurements of the Pep2-2 derivatives (Fig. 1C) and for the truncated Pep2-8 peptides (Table 1), we used the peptides in a concentration range of 0-250 μM and the Pep2-2-displaying phage at subsaturating concentrations. |
| Affinity data for this assay | |
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