Recombinant human PCSK9 (Proprotein convertase subtilisin/kexin type 9), encompassing amino acids 31-692 (end). The protein contains mutation of interest D374T, and a C-terminal His-tag (6xHis) followed by an Avi-Tag™. The recombinant protein was enzymatically biotinylated using the Avi-Tag™ and affinity purified. HiP™ indicates a high purity protein (≥90% pure) and less than 10% aggregation as measured by gel filtration.
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Synonyms
Proprotein convertase subtilisin/kexin type 9, FH3, HCHOLA3, PC9
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Product Data Gallery
4-20% SDS-PAGE Coomassie Staining
PCSK9 (D374T): LDLR Activity
Biotin-Avidin Pulldown
Gel Filtration Trace
PCSK9(D374T)-LDLR inhibition curve using a chemiluminescent assay (#78326-1). Representative data only; data may vary by lot.
40 mM Tris-HCl, pH 8.0, 110 mM NaCl, 2.2 mM KCl, and 20% glycerol
MW
Pro: 14 kDa + glycans; Mature: 60 kDa + glycans
Amino Acids
31-692(end)
Glycosylation
This protein runs at a higher MW by SDS-PAGE due to glycosylation.
Aggregation
<10%
Genbank #
NM_174936
UniProt #
Q8NBP7
Tag(s)
C-terminal His, Avi-tags
Label
This protein is enzymatically biotinylated using Avi-Tag™ technology. Biotinylation confirmed to be ≥90%.
For more information on enzymatic biotinylation, please see our Tech Note.
Background
PCSK9 is a crucial player in the regulation of plasma cholesterol homeostasis. It binds to the ectodomain of hepatic low-density lipid receptor family members: low density lipoprotein receptor (LDLR), very low density lipoprotein receptor (VLDLR), apolipoprotein E receptor (LRP1/APOER) and apolipoprotein receptor 2 (LRP8/APOER2), and promotes their degradation. PCSK9 acts via a non-proteolytic mechanism to enhance the degradation of the hepatic LDLR through a clathrin LDLRAP1/ARH-mediated pathway. May prevent the recycling of LDLR from endosomes to the cell surface or direct it to lysosomes for degradation. The D374T mutation results in higher affinity of PCSK9 for LDLR.
Storage/Stability
At least 6 months at –80°C.
Assay Conditions
Assay was done according to the PCSK9(D374T)-LDLR TR-FRET Assay Kit (BPS Bioscience #72011) with PCSK9 (D374T) titrated from 160 ng-2.5 ng/rxn.
Instructions for Use
Thaw on ice and gently mix prior to use. DO NOT VORTEX. Perform a quick spin before opening. Aliquot into small volumes and flash freeze for long term storage. Avoid multiple freeze/thaw cycles.
Applications
Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
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