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Applications: Great for screening small molecules and antibodies that inhibit the binding of PCSK9 to LDLR ectodomain.
Background: PCSK9 is a crucial player in the regulation of plasma cholesterol homeostasis. It binds to the ectodomain of hepatic low-density lipid receptor family members: low density lipoprotein receptor (LDLR), very low density lipoprotein receptor (VLDLR), apolipoprotein E receptor (LRP1/APOER) and apolipoprotein receptor 2 (LRP8/APOER2), and promotes their degradation. Acts via a non-proteolytic mechanism to enhance the degradation of the hepatic LDLR through a clathrin LDLRAP1/ARH-mediated pathway. May prevent the recycling of LDLR from endosomes to the cell surface or direct it to lysosomes for degradation.
Contraindications: DMSO >1%, strong acids or bases, ionic detergents, high salt
Description: The PCSK9[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 by streptavidin-HRP. Only a few simple steps on a microtiter plate are required for the assay. First, LDLR ectodomain is coated on a 96-well plate. Next, PCSK9 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: Mixed storage conditions. Stable at least 12 months from date of receipt, when stored as directed
Supplied As: The PCSK9[Biotinylated]-LDLR Binding Assay Kit comes in a convenient 96-well format, with biotin-labeled PCSK9, purified LDLR ectodomain, streptavidin labeled HRP, and assay buffer for 96 binding reactions.
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Chan, J.C. et al. (2009). Proc. Natl Acad. Sci. USA, 106, 9820-9825.
2. Liang, H., et al. (2012) J. Pharmacol. Exp. Ther. 340 2289-236
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