膜杰作
Star Staining
分子别名（Synonym）
PCSK9,FH3,HCHOLA3,LDLCQ1,NARC1,PC9
表达区间及表达系统（Source）
MABSol® Biotinylated Human PCSK9 (D374Y), Avitag,His Tag (PCY-H82E7) is expressed from human HEK293 cells. It contains AA Gln 31 - Gln 692 (Accession # Q8NBP7-1 (D374Y)).
Predicted N-terminus: Gln 31
Request for sequence
蛋白结构（Molecular Characterization）
This protein carries an Avi tag (Avitag™) at the C-terminus, followed by a polyhistidine tag. 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 59.9 kDa respectively. The protein migrates as 17 kDa and 60-65 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation. The D374Y mutation results in higher affinity of PCSK9 for LDLR.
标记（Labeling）
Biotinylation of this product is performed using Avitag™ technology. Briefly, the single lysine residue in the Avitag is enzymatically labeled with biotin.
蛋白标记度（Protein Ratio）
Passed as determined by the HABA assay / binding ELISA.
内毒素（Endotoxin）
Less than 1.0 EU per μg by the LAL method.
纯度（Purity）
>95% as determined by SDS-PAGE.
制剂（Formulation）
Lyophilized from 0.22 μm filtered solution in PBS, pH7.4 with trehalose as protectant.
Contact us for customized product form or formulation.
重构方法（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.
存储（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.
电泳（SDS-PAGE）
Biotinylated Human PCSK9 (D374Y), Avitag,His Tag on SDS-PAGE under reducing (R) condition. The gel was stained with Coomassie Blue. The purity of the protein is greater than 95%.
活性（Bioactivity）-ELISA
Immobilized Human LDL R, His Tag (Cat. No. LDR-H5224) at 2 μg/mL (100 μL/well) can bind Biotinylated Human PCSK9 (D374Y), Avitag,His Tag (Cat. No. PCY-H82E7) with a linear range of 0.2-8 ng/mL (QC tested).
Protocol
Immobilized Biotinylated Human PCSK9 (D374Y), Avitag,His Tag (Cat. No. PCY-H82E7) at 1 μg/mL (100 μL/well) on streptavidin (Cat. No. STN-N5116) precoated (0.5 μg/well) plate can bind Evolocumab with a linear range of 0.1-1 ng/mL (Routinely tested).
Protocol
背景（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).
