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Your Position: Home > Inhibitory Kit > PCSK9 > EP-103

PCSK9 [Biotinylated] : LDL R Inhibitor Screening ELISA Assay Pair

For research use only.
Materials Provided
IDComponentsSize (96 tests)Size (480 tests)
A006-214Human LDL R35 μg160 μg
A007-214Biotinylated Human PCSK910 μg10 μg
A003-214Streptavidin-HRP10 μg10 μg
PC9-NA003Anti-PCSK9 Neutralizing Antibody40 μg200 μg
  • Background
    PCSK9 is a crucial protein in the regulation of plasma cholesterol homeostasis. It binds low density lipoprotein receptor (LDL R) to enhance the degradation of LDL R. Therefore, inhibition of PCSK9 has been considered a promising strategy to prevent the receptor from being degraded and promote removal of LDL cholesterol from circulation.
  • Application

    This pair is useful for screening for inhibitors of human PCSK9 binding to human LDL R.

  • Reconstitution
    See Certificate of Analysis for details of reconstitution instruction and specific concentration.
  • Storage

    Upon receipt, please store all items at -70℃.

    After reconstitution, the stock solution should be kept at -70℃.

    It is recommended not to freeze thaw more than 3 times.

    This product is stable after storage at:

    • Room temperature (RT) for 1 month in lyophilized state;

    • -20℃ for 1 year in lyophilized state;

    • -70℃ for 1 year under sterile conditions after reconstitution

  • Assay Principles

    This inhibitor screening ELISA pair is designed to facilitate the identification and characterization of new PCSK9 pathway inhibitors. This assay employs a simple colorimetric ELISA platform, which measures the binding between immobilized human LDL R and in-house developed biotinylated PCSK9 protein. This product is uniquely suitable for rapid high-throughput screening of putative PCSK9 inhibitors. Briefly, we provide you with a biotinylated human PCSK9 protein, a human LDL R protein, an anti-PCSK9 neutralizing antibody (as method verified Reference), and Streptavidin-HRP reagent. Your experiment will include 4 simple steps:

    • a) Coat the plate with human LDL R.

    • b) Add your molecule of interest to the plates.

    • c) Add human PCSK9-Biotin to the plates.

    • d) Add Streptavidin-HRP followed by TMB or other colorimetric HRP substrate.
      Finally, the ability of your compound to inhibit PCSK9: LDL R binding will be determined by comparing OD readings among different experimental groups.

Typical Data

Binding of Biotinylated Human PCSK9 to Immobilized Human LDL R in a Functional ELISA Assay.

Immobilized human LDL R protein at 3 μg/mL (100 μL/well) can bind biotinylated human PCSK9 with a linear range of 0.000625-0.02 μg/mL when detected by Streptavidin-HRP. Background was subtracted from data points before curve fitting.

Inhibition of PCSK9 [Biotinylated] : LDL R Binding by Anti-PCSK9 Neutralizing Antibody.

Serial dilutions of anti-PCSK9 neutralizing antibody (Catalog # PC9-NA003) (1:2 serial dilutions, from 20 μg/mL to 0.04 μg/mL) were added into PCSK9: biotinylated LDL R binding reactions. The assay was performed according to the above described protocol. Background was subtracted from data points prior to log transformation and curve fitting.

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