PCSK9(His)-LDLR Binding Assay Kit

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Catalog #

78813

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Description

The PCSK9(His)-LDLR Binding Assay Kit is an ELISA-type 96-well format assay designed for screening and profiling purposes. Moreover, two pre-formulated assay buffers are supplied to validate PCSK9-LDLR binding affinity in either neutral or acidic binding conditions. The assay takes advantage of the high sensitivity of detection of His-tag PCSK9 by Anti-His HRP-antibody. First, the LDLR ectodomain is coated on a 96-well plate. Next, PCSK9 is incubated with LDLR on the plate. Finally, the plate is treated with Anti-His HRP-antibody followed by addition of an HRP substrate to produce chemiluminescence, measured using a chemiluminescence reader.

Figure 1: Illustration of the ELISA-based PCSK9(His)-LDLR Binding Assay Kit.

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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

Chemiluminescent

Supplied As

The Assay Kit contains His-tag PCSK9, purified LDLR ectodomain and Anti-His-tag Horseradish Peroxidase (HRP)-conjugated antibody in sufficient amount for 100 binding reactions.

Materials Required But Not Supplied

PBS (Phosphate Buffer Saline)
Luminometer or microplate reader capable of reading chemiluminescence
Rotating or rocker platform

Format

Catalog #	Name	Amount	Storage
71204	PCSK9(His)*	10 µg	-80°C
71205	LDLR*	10 µg	-80°C
	Anti-His HRP-D Antibody-D	10 µl	-80°C
33298	3x PL-01 Assay Buffer	50 ml	-20°C
79727	3x PL-02 Assay Buffer	50 ml	-20°C
79728	Blocking Buffer 2	50 ml	+4°C
79670	ELISA ECL Substrate A (translucent bottle)	6 ml	Room Temp
79670	ELISA ECL Substrate B (brown bottle)	6 ml	Room Temp
79699	White 96-well microplate	1	Room Temp

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

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 molecules and antibodies that inhibit the binding of PCSK9 to LDLR ectodomain.

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]

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 solution in buffer and using 5 µl per well.

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(His)-LDLR Binding Assay Kit

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