Human 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 31
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.
Less than 1.0 EU per μg by the LAL method.
>92% as determined by SDS-PAGE.
Lyophilized from 0.22 μm filtered solution in PBS, pH7.4. Normally trehalose is added as protectant before lyophilization.
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Please see Certificate of Analysis for specific instructions.
For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.
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, Fc Tag (Cat. No. LDR-H5254) at 2 μg/mL (100 μL/well) can bind Human PCSK9 (D374Y), His Tag (Cat. No. PCY-H5225) with a linear range of 20-78 ng/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.
Proprotein 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.