- SynonymPCSK9,FH3,HCHOLA3,LDLCQ1,NARC1,PC9
- SourceHuman PCSK9 (D374Y), His Tag (PCY-H5225) is expressed from human 293 cells (HEK293). It contains AA Gln 31 - Gln 692 (Accession # Q8NBP7-1(D374Y)).Predicted N-terminus: Gln 31Request for sequence
- Molecular Characterization
This protein carries a polyhistidine tag at the C-terminus. This protein undergoes autocatalytic cleavage to release the pro-peptide and mature chain. The pro-peptide and mature chain are associated through non‑covalent interactions and with a calculated MW of 13.8 kDa and 58.2 kDa respectively. The protein migrates as 17 kDa and 66 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation. The D374Y mutation results in higher affinity of PCSK9 for LDLR.
- EndotoxinLess than 1.0 EU per μg by the LAL method.
- Purity
>92% as determined by SDS-PAGE.
- Formulation
Lyophilized from 0.22 μm filtered solution in PBS, pH7.4. Normally trehalose is added as protectant before lyophilization.
Contact us for customized product form or formulation.
- Reconstitution
Please see Certificate of Analysis for specific instructions.
For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.
- Storage
For long term storage, the product should be stored at lyophilized state at -20°C or lower.
Please avoid repeated freeze-thaw cycles.
This product is stable after storage at:
- -20°C to -70°C for 12 months in lyophilized state;
- -70°C for 3 months under sterile conditions after reconstitution.
Human PCSK9 (D374Y), His Tag on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 92%.
Immobilized Human LDL R, His Tag (Cat. No. LDR-H5224) at 10 μg/mL (100 μL/well) can bind Human PCSK9 (D374Y), His Tag (Cat. No. PCY-H5225) with a linear range of 0.004-0.063 μg/mL (QC tested).
Loaded Human LDL R, Fc Tag (Cat. No. LDR-H5254) on Protein A Biosensor, can bind Human PCSK9 (D374Y), His Tag (Cat. No. PCY-H5225) with an affinity constant of 0.329 nM as determined in BLI assay (ForteBio Octet Red96e) (Routinely tested).
FACS analysis shows that Human PCSK9 (D374Y), His Tag (Cat. No. PCY-H5225) inhibits LDL uptake in HepG2 cells. The EC50 for this effect is 0.0689-0.3049 μg/mL.
FACS analysis shows that the effect of Human PCSK9 (D374Y), His Tag (Cat. No. PCY-H5225) inhibiting LDL uptake in HepG2 cells was neutralized by Anti-Human PCSK9 antibody. The concentration of PCSK9 used is 5 μg/mL. The EC50 for Anti-Human PCSK9 antibody is 6.816-12.67 μg/mL.
- BackgroundProprotein convertase subtilisin/kexin type 9 (PCSK9), is an enzyme which in humans is encoded by the PCSK9 gene. This gene encodes a proprotein convertase belonging to the proteinase K subfamily of the secretory subtilase family. This protein plays a major regulatory role in cholesterol homeostasis. PCSK9 binds to the epidermal growth factor-like repeat A (EGF-A) domain of the low-density lipoprotein receptor (LDLR), inducing LDLR degradation. PCSK9 may also have a role in the differentiation of cortical neurons. Mutations in this gene have been associated with a rare form of autosomal dominant familial hypercholesterolemia (HCHOLA3).Especially,the D374Y mutation results in higher affinity of PCSK9 for LDLR.
- References
- (1)Seidah NG, et al., 2003, Proc. Natl. Acad. Sci. U.S.A. 100 (3): 928–33.
- (2)Abifadel, M. et al., 2003, Nat. Genet. 34: 154-156.
- (3)Dubuc G. et al., 2004, Arterioscler. Thromb. Vasc. Biol. 24 (8): 1454–9.
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