PCSK9(D374Y) [Biotinylated]-LDLR Binding Assay Kit

Catalog #

78866

$805 *

Size: 96 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(D374Y) [Biotinylated]-LDLR Binding Assay Kit is designed for screening and profiling purposes. The kit comes in a convenient 96-well format and contains enough biotin-labeled mutant protein PCSK9(D374Y), purified LDLR ectodomain, and HRP-conjugated streptavidin for 100 reactions. Moreover, two pre-formulated assay buffers are supplied to validate PCSK9-LDLR binding in either neutral or acidic binding conditions.

This assay takes advantage of the high sensitivity of detection of biotin-labeled PCSK9(D374Y) by streptavidin-HRP. First, a 96-well plate is coated with LDLR ectodomain. PCSK9(D374Y) is then incubated with LDLR. Finally, the plate is treated with streptavidin-HRP followed by addition of an HRP substrate to produce chemiluminescence, which is measured using a chemiluminescence reader.

Need us to run inhibitor screens or profile your compounds against PCSK9(D374Y)-LDLR? Check out our PCSK9 Screening Services.

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Product Data Gallery

PCSK9-LDLR binding activity under acidic or neutral conditions.

Product Info

Storage and Usage

Citations

Assay Kit Format

Chemiluminescent

Mutation

D374Y

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
101725	PCSK9(D374Y), Biotin-Labeled*	10 µg	-80°C
71205	LDLR, FLAG-Tag*	10 µg	-20°C
79742	Streptavidin-HRP	10 µl	-20°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 Substrates A and B (2 components)	6 ml each	Room Temp
79699	96-well plate, white	1	+4°C

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

Background

PCSK9 (Proprotein convertase subtilisin/kexin type 9) functions as a negative regulator of the hepatic low-density lipoprotein receptors (LDLRs) by binding to the LDLR ectodomain. The D374Y mutation is associated with severe 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 LDLR.

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(D374Y) to the LDLR ectodomain in high-throughput screening applications (HTS).

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 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-Related

Regulatory Status

Research Use Only