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Application: Screen small molecules and antibodies that inhibit the binding of PCSK9 to LDLR ectodomain.
Background: PCSK9 (Proprotein convertase subtilisin/kexin type 9) functions as a negative regulator of hepatic low-density lipoprotein receptors (LDLRs) and therefore is a critical regulator of cholesterol metabolism. It is an endopeptidase that binds to the EGFR-like ectodomain of LDLR, leading to LDLR degradation, which in turn, results in increased circulating LDL. Inhibiting the PCSK9-LDLR interaction is an increasingly desirable therapeutic approach for lowering LDL-cholesterol levels to replace or supplement statins. New therapies are critical for addressing atherosclerosis, stroke, heart disease, and other cardiovascular disorders.
Contraindications: PCSK9(His)-LDLR Binding Assay Kit is compatible with up to 1% final DMSO concentration. We recommend preparing the inhibitor in no higher than 10% DMSO solution in buffer and using 5 µl per well.
Description: The PCSK9(His)-LDLR Binding Assay Kit is an ELISA-type 96-well format assay designed for screening and profiling purposes. Moreover, two pre-formulated assay buffers are supplied to validate PCSK9-LDLR binding affinity in either neutral or acidic binding conditions. The assay takes advantage of the high sensitivity of detection of His-tag PCSK9 by Anti-His HRP-antibody. First, the LDLR ectodomain is coated on a 96-well plate. Next, PCSK9 is incubated with LDLR on the plate. Finally, the plate is treated with Anti-His HRP-antibody followed by addition of an HRP substrate to produce chemiluminescence, measured using a chemiluminescence reader. Figure 1: Illustration of the ELISA-based PCSK9(His)-LDLR Binding Assay Kit.
Storage Stability: This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.
Supplied As: The Assay Kit contains His-tag PCSK9, purified LDLR ectodomain and Anti-His-tag Horseradish Peroxidase (HRP)-conjugated antibody in sufficient amount for 100 binding reactions.
Uniprot: PCSK9: Q8NBP7; LDLR: P01130
Warnings: Avoid freeze/thaw cycles
Biosafety Level: Not applicable (BSL-1)