DESCRIPTION

• Notes: Troubleshooting Guide: Visit bpsbioscience.com/assay-kits-faq for detailed troubleshooting instructions. For all further questions, please email support@bpsbioscience.com. TrademarksAlphaScreen® and AlphaLISA® are registered trademarks of PerkinElmer, Inc.
• Uniprot: PCSK9: Q8NBP7; LDLR: P01130
• Shiptemp: -80°C (dry ice)
• Synonyms: Low-density lipoprotein receptor, LDL receptor, Proprotein convertase subtilisin/kexin type 9, Neural apoptosis-regulated convertase 1, NARC-1, Proprotein convertase 9, PC9, Subtilisin/kexin-like protease PC9, NARC1, PSEC0052
• Warnings: Avoid freeze/thaw cycles
• Category: Cholesterol/Assay Kit
• 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.
• Description: The PCSK9-LDLR Homogeneous Assay Kit is designed for screening and profiling purposes. Two pre-formulated assay buffers are supplied with this kit to validate PCSK9-LDLR binding affinity in either neutral or acidic binding conditions. With this kit, two simple steps are required for binding measurement. First, PCSK9 enzyme is incubated with LDLR in the preferred buffer for one hour. Next, donor and acceptor beads are added, followed by reading the Alpha-counts. Figure 1: Illustration of the assay principle. The FLAG-tag of LDLR binds to the anti-FLAG acceptor beads, while the His-tag on PCSK9 binds to Nickel Chelate donor beads, bringing the acceptor and donor beads in close proximity. Upon excitation of the donor bead a singlet oxygen is generated by the donor bead, which excites the acceptor bead and emits light proportionally to the level of interaction. AlphaLISA™ immunoassays are a no-wash alternative to ELISA immunoassays. These assays are robust, ideal for a minimal hands-on approach, and highly amenable to high-throughput applications.
• Supplied As: The PCSK9-LDLR Homogeneous Assay Kit comes in a no-wash AlphaLISA® format, with sufficient PCSK9 and LDLR for 384 enzyme reactions.
• Unspsc Code: 41121811
• Unspsc Name: Laboratory microchemistry kits
• Applications: Screen small molecule inhibitors in high throughput (HTS) applications.
• Product Type: Assay Kit
• Biosafety Level: Not applicable (BSL-1)
• Related Products: 72010, 72011, 71211, 71305
• Contraindications: Green and blue dyes that absorb light in the AlphaScreen® signal emission range (520-620 nm), such as Trypan Blue, interfere with the assay. Avoid using potent singlet oxygen quenchers such as sodium azide (NaN3) or metal ions (Fe2+, Fe3+, Cu2+, Zn2+ and Ni2+). The presence of >1% RPMI 1640 culture medium leads to a signal reduction due to the presence of excess biotin and iron in this medium. MEM, which lacks these components, does not affect AlphaScreen® assays.
• Storage Stability: This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.
• Scientific Category: Cholesterol