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Application: Study enzyme kinetics and screen small molecular inhibitors in high throughput (HTS) applications.
Background: PCSK9 (Proprotein convertase subtilisin/kexin type 9) functions as a negative regulator of hepatic low-density lipoprotein receptors (LDLRs) by binding to the LDLR ectodomain. The D374T mutation is associated with 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 the LDLR.
Description: The PCSK9(D374T) [Biotinylated]-LDLR Binding Assay Kit is designed for screening and profiling purposes. The key to this kit is the high sensitivity of detection of biotin-labeled PCSK9(D374T) by streptavidin-HRP. Only a few steps on a microtiter plate are required for the assay. First, LDLR ectodomain is coated on a 96-well plate. Next, PCSK9(D374T) is incubated with LDLR on the plate. Finally, the plate is treated with streptavidin-HRP followed by addition of an HRP substrate to produce chemiluminescence, which can then be 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.
Supplied As: The kit comes in a convenient 96-well format, with biotin-labeled mutant protein PCSK9(D374T), purified LDLR ectodomain, streptavidin labeled HRP, and assay buffer for 100 binding reactions.
Uniprot: PCSK9: Q8NBP7; LDLR: P01130
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Chan, J.C. et al. (2009). Natl Acad. Sci. USA, 106, 9820-9825.
2. Liang, H., et al. (2012) Pharmacol. Exp. Ther. 340, 2289-236.