组分(Materials Provided)
ID | Components | Size |
A006-214 | Human LDL R | 35μg |
A007-214 | Biotinylated Human PCSK9 | 10μg |
A003-214 | Streptavidin-HRP | 10μg |
PC9-NA003 | Anti-PCSK9 Neutralizing Antibody | 40μg |
ID | Components | Size |
A006-214 | Human LDL R | 160μg |
A007-214 | Biotinylated Human PCSK9 | 10μg |
A003-214 | Streptavidin-HRP | 10μg |
PC9-NA003 | Anti-PCSK9 Neutralizing Antibody | 200μg |
背景(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.
应用说明(Application)
This pair is useful for screening for inhibitors of human PCSK9 binding to human LDL R.
It is for research use only.
重构方法(Reconstitution)
See Certificate of Analysis for details of reconstitution instruction and specific concentration.
存储(Storage)
Upon receipt, please store all items at -70°C.
After reconstitution, the stock solution should be kept at -70°C.
原理(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:
1. Coat the plate with human LDL R.
2. Add your molecule of interest to the plates.
3. Add human PCSK9-Biotin to the plates.
4. 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.
典型数据-Typical Data Please refer to Ds document for the assay protocol.
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 or 133.33 nM to 0.26 nM) were added into Biotinylated PCSK9: LDL R binding reactions. Background was subtracted from data points prior to log transformation and curve fitting (QC tested).