PCSK9-LDLR Homogeneous Assay Kit

Catalog #

78812

$1,150 *

Size: 384 reactions

Qty

Bulk/Custom

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*US Pricing only. For international pricing, please contact your local distributor.

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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.

Need us to run inhibitor screens or profile your compounds against PCSK9-LDLR? Check out our PCSK9 Screening Services.

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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

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Product Data Gallery

PCSK9-LDLR binding activity in neutral or acidic buffer

Product Info

Storage and Usage

Citations

Assay Kit Format

AlphaLISA®

Supplied As

The PCSK9-LDLR Homogeneous Assay Kit comes in a no-wash AlphaLISA® format, with sufficient PCSK9 and LDLR for 384 enzyme reactions.

Materials Required But Not Supplied

AlphaLISA® anti-FLAG acceptor beads, 5 mg/ml (PerkinElmer #AL112C)
AlphaScreen® Nickel Chelate donor beads, 5 mg/ml (PerkinElmer #AS101D)
Optiplate-384 (PerkinElmer #6007290)

Format

Catalog #	Name	Amount	Storage
71204	PCSK9, His-Tag Recombinant*	2 x 10 µg	-80°C
71205	LDLR, FLAG-Tag Recombinant*	2 x 10 µg	-20°C
33298	3x PL-01 Assay Buffer (pH 6)	4 ml	-20°C
79727	3x PL-02 Assay Buffer (pH 7.4)	4 ml	-20°C

* The concentration of protein is lot-specific and will be indicated on the tube containing the protein

UniProt #

PCSK9: Q8NBP7; LDLR: P01130

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.

Storage/Stability

This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.

Applications

Screen small molecule inhibitors in high throughput (HTS) applications.

Shipping Temperature

-80°C

Notes

Troubleshooting Guide
Visit bpsbioscience.com/assay-kits-faq for detailed troubleshooting instructions. For all further questions, please email [email protected].

Trademarks
AlphaScreen® and AlphaLISA® are registered trademarks of PerkinElmer, Inc.

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.

Warnings

Avoid freeze/thaw cycles

Disclaimers and Limitations

BPS Bioscience assay kits are validated using the listed components according to our specific protocol. Any deviations to kit components or protocols, such as using different proteins, cell/tissue lysates, or buffers (including using the same component from other commercial sources) will void any warranties on the performance of the assay kit and is not recommended.

End-users should immediately report any issues to [email protected] upon receipt of the kit, such as missing components, damaged vials, no dry ice, etc.

Scientific Category

Cholesterol

Regulatory Status

Research Use Only

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PCSK9-LDLR Homogeneous Assay Kit

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