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Your Position: Home > Protein > PCSK9 > PCY-H5225

Human PCSK9 (D374Y) Protein, His Tag

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  • Synonym
    PCSK9,FH3,HCHOLA3,LDLCQ1,NARC1,PC9
  • Source
    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
  • Molecular Characterization
    PCSK9 Structure

    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.

  • Endotoxin
    Less than 1.0 EU per μg by the LAL method.
  • Purity

    >90% as determined by SDS-PAGE.

  • Formulation

    Lyophilized from 0.22 μm filtered solution in PBS, pH7.4 with trehalose as protectant.

    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:

    1. -20°C to -70°C for 12 months in lyophilized state;
    2. -70°C for 3 months under sterile conditions after reconstitution.
SDS-PAGE
PCSK9 SDS-PAGE

Human PCSK9 (D374Y), His Tag on SDS-PAGE under reducing (R) condition. The gel was stained with Coomassie Blue. The purity of the protein is greater than 90%.

Bioactivity-ELISA
 PCSK9 ELISA

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).

 PCSK9 ELISA

Immobilized Human PCSK9 (D374Y), His Tag (Cat. No. PCY-H5225) at 1 μg/mL (100 μL/well) on Monoclonal Anti-His Tag Antibody, Mouse IgG1 (AY63) (Cat. No. HIS-AY63) precoated (0.1 μg/well) plate can bind Evolocumab with a linear range of 0.1-4 ng/mL (Routinely tested).

Bioactivity-BLI
 PCSK9 BLI

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).

Bioactivity-FACS
 PCSK9 FACS

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.

 PCSK9 FACS

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.

  • Background
    Proprotein convertase subtilisin/kexin type 9 (PCSK9) is also known as NARC1 (neural apoptosis regulated convertase), is a newly identified subtilase belonging to the peptidase S8 subfamily. Mouse PCSK9 is synthesized as a soluble zymogen, and undergoes autocatalytic intramolecular processing in the endoplasmic reticulum, resulting in the cleavage of its propeptide that remains associated with the secreted active enzyme with a broad alkaline pH optimum. 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).
  • Clinical and Translational Updates

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