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Applications: Screen small molecules and antibodies that inhibit the binding of PCSK9(D374Y) to the LDLR ectodomain in high-throughput screening applications (HTS).
Background: PCSK9 (Proprotein convertase subtilisin/kexin type 9) functions as a negative regulator of the hepatic low-density lipoprotein receptors (LDLRs) by binding to the LDLR ectodomain. The D374Y mutation is associated with severe hypercholesterolemia; this form of PCSK9 is more potent at decreasing LDL uptake than wild-type PCSK9, most likely by increasing the binding affinity of PCSK9 for LDLR.
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 and using 5 µl per well.
Description: The PCSK9(D374Y) [Biotinylated]-LDLR Binding Assay Kit is designed for screening and profiling purposes. The kit comes in a convenient 96-well format and contains enough biotin-labeled mutant protein PCSK9(D374Y), purified LDLR ectodomain, and HRP-conjugated streptavidin for 100 reactions. Moreover, two pre-formulated assay buffers are supplied to validate PCSK9-LDLR binding in either neutral or acidic binding conditions.This assay takes advantage of the high sensitivity of detection of biotin-labeled PCSK9(D374Y) by streptavidin-HRP. First, a 96-well plate is coated with LDLR ectodomain. PCSK9(D374Y) is then incubated with LDLR. Finally, the plate is treated with streptavidin-HRP followed by addition of an HRP substrate to produce chemiluminescence, which is measured using a chemiluminescence reader.
Storage Stability: This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.
Warnings: Avoid freeze/thaw cycles
Biosafety Level: Not applicable (BSL-1)
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