PCSK9(D374T) [Biotinylated]-LDLR Binding Assay Kit

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Catalog #

78326

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Description

The PCSK9(D374T) [Biotinylated]-LDLR Binding Assay Kit is designed for screening and profiling purposes. This assay takes advantage of the high sensitivity of detection of biotin-labeled PCSK9(D374T) by streptavidin-HRP. Only a few steps are required for the assay. First, LDLR ectodomain is coated on a 384-well plate. Next, PCSK9(D374T) 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.

Need us to run inhibitor screens or profile your compounds against PCSK9(D374T)-LDLR? Check out our PCSK9 Screening Services.

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Synonyms

Biotinylated PCSK9, FH3, HCHOLA3, PC9, NARC1, Neural Apoptosis Regulated Convertase 1, FH, FHC, LDLR, LDLCQ2, low density lipoprotein receptor

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Product Data Gallery

PCSK9 (D374T) Binding Activity

LDLR: PCSK9(D374T) Binding Activity

Product Info

Storage and Usage

Citations

Assay Kit Format

Chemiluminescent

Mutation

D374T

Supplied As

The kit comes in a convenient 96 or 384-well format, with biotin-labeled mutant protein PCSK9(D374T), purified LDLR ectodomain, streptavidin-labeled HRP, and assay buffer for 100 or 384 reactions.

Materials Required But Not Supplied

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

Format

96 reactions

Catalog #	Name	Amount	Storage
71211	PCSK9(D374T), Biotin-Labeled*	10 µg	-80°C
71205	LDLR*	10 µg	-20°C
79742	Streptavidin-HRP	10 µl	-20°C
33298	3x PL-01 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

384 reactions

Catalog #	Name	Amount	Storage
71211	PCSK9(D374T), Biotin-Labeled*	2 x 10 µg	-80°C	*Avoid multiple freeze/ thaw cycles*
71205	LDLR, FLAG-Tag*	2 x 10 µg	-80°C
79742	Streptavidin-HRP	2 x 10 µl	4°C
33298	3x PL-01 Assay Buffer	2 x 50 ml	-20°C
79728	Blocking Buffer 2	2 x 50 ml	4°C
79670	ELISA ECL Substrate A (translucent bottle)	2 x 6 ml	Room Temp
79670	ELISA ECL Substrate B (brown bottle)	2 x 6 ml	Room Temp
78188	384-well plate	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) by binding to the LDLR ectodomain. The D374T mutation is associated with 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 the LDLR.

References

1. Chan, J.C. et al. (2009). Natl Acad. Sci. USA, 106, 9820-9825.
2. Liang, H., et al. (2012) Pharmacol. Exp. Ther. 340, 2289-236.

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

Study enzyme kinetics and screen small molecular 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]

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