Human PCSK9 ELISA Kit EZ-SetTM (DIY Antibody Pairs)

Proprotein convertase subtilisin/kexin type 9, also known as PCSK9, is an enzyme that 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. By genomic sequence analysis, the PCSK9 gene was mapped to chromosome 1p32. 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. Reduced LDLR levels result in decreased metabolism of low-density lipoproteins (LDL), which could lead to hypercholesterolemia. PCSK9 may also have a role in the differentiation of cortical neurons.

Convertase subtilisin/kexin type 9 preproprotein; FH3; HCHOLA3; Hypercholesterolemia autosomal dominant 3; LDLCQ1; NARC 1; NARC-1; NARC1; Neural apoptosis regulated convertase 1; Neural apoptosis-regulated convertase 1; PC 9; PC9; PCSK 9; PCSK9; PCSK9_HUMAN; Proprotein convertase 9; Proprotein convertase PC9; Proprotein convertase subtilisin/kexin type 9; PSEC0052; Subtilisin/kexin like protease PC9; Subtilisin/kexin-like protease PC9

Amino Acid Metabolism, Amino Acids, Cancer, Cardiovascular, Cell Biology, Cholesterol Metabolism, Intracellular, Lipid and Lipoprotein Metabolism, Lipid Metabolism, Lipids/Lipoproteins, Metabolic Signaling Pathways, Metabolism, Neural Stem Cells, Pathways and Processes, Proteolysis/Ubiquitin, Proteolytic Enzymes, Signal Transduction, Stem Cells

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 229 1991 5404 215 1927 5944 Standard deviation 16.03 125.43 243.18 17.63 132.96 285.31 CV (%) 7% 6.3% 4.5% 8.2% 6.9% 4.8% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 229 1991 5404 215 1927 5944 Standard deviation 16.03 125.43 243.18 17.63 132.96 285.31 CV (%) 7% 6.3% 4.5% 8.2% 6.9% 4.8% Intra-Assay Precision Inter-Assay Precision Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 n 16 16 16 24 24 24 Mean (pg/ml) 229 1991 5404 215 1927 5944 Mean (pg/ml) 229 1991 5404 215 1927 5944 Standard deviation 16.03 125.43 243.18 17.63 132.96 285.31 Standard deviation 16.03 125.43 243.18 17.63 132.96 285.31 CV (%) 7% 6.3% 4.5% 8.2% 6.9% 4.8% CV (%) 7% 6.3% 4.5% 8.2% 6.9% 4.8%

Crucial player in the regulation of plasma cholesterol homeostasis. Binds to low-density lipid receptor family members: low density lipoprotein receptor (LDLR), very low density lipoprotein receptor (VLDLR), apolipoprotein E receptor (LRP1/APOER) and apolipoprotein receptor 2 (LRP8/APOER2), and promotes their degradation in intracellular acidic compartments. Acts via a non-proteolytic mechanism to enhance the degradation of the hepatic LDLR through a clathrin LDLRAP1/ARH-mediated pathway. May prevent the recycling of LDLR from endosomes to the cell surface or it to lysosomes for degradation. Can induce ubiquitination of LDLR leading to its subsequent degradation. Inhibits intracellular degradation of APOB via the autophagosome/lysosome pathway in a LDLR-independent manner. Involved in the disposal of non-acetylated intermediates of BACE1 in the early secretory pathway. Inhibits epithelial Na+ channel (ENaC) -mediated Na+ absorption by reducing ENaC surface expression primarily by increasing its proteasomal degradation. Regulates neuronal apoptosis via modulation of LRP8/APOER2 levels and related anti-apoptotic signaling pathways.

Catalog number Description Quantity Storage of opened/reconstituted material EZ1147-CA Rabbit anti- human PCSK9 polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ1147-DA Biotinylated goat anti- human PCSK9 polyclonal antibody (Detection Antibody) 500 μl, 28 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK1147-ST Lyophilized recombinant human PCSK9 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. Catalog number Description Quantity Storage of opened/reconstituted material EZ1147-CA Rabbit anti- human PCSK9 polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ1147-DA Biotinylated goat anti- human PCSK9 polyclonal antibody (Detection Antibody) 500 μl, 28 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK1147-ST Lyophilized recombinant human PCSK9 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. Catalog number Description Quantity Storage of opened/reconstituted material Catalog number Description Quantity Storage of opened/reconstituted material EZ1147-CA Rabbit anti- human PCSK9 polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ1147-CA Rabbit anti- human PCSK9 polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ1147-DA Biotinylated goat anti- human PCSK9 polyclonal antibody (Detection Antibody) 500 μl, 28 μg/mL (recommended dilution 1:100) EZ1147-DA Biotinylated goat anti- human PCSK9 polyclonal antibody (Detection Antibody) 500 μl, 28 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK1147-ST Lyophilized recombinant human PCSK9 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. EK1147-ST Lyophilized recombinant human PCSK9 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit.

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.058 0.112 0.14 0.235 0.391 0.73 1.314 2.215 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.058 0.112 0.14 0.235 0.391 0.73 1.314 2.215 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.058 0.112 0.14 0.235 0.391 0.73 1.314 2.215 O.D. 0.058 0.112 0.14 0.235 0.391 0.73 1.314 2.215

1. Microplate reader in standard size. 2. Automated plate washer. 3. Incubator. 4. Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection. 5. Clean tubes and Eppendorf tubes. 6. 96 well microplate (Cat# AR1100) 7. Plate Sealers. 8. Capture Antibody Diluent: PBS. 9. Reagent Diluent: 1% BSA in PBS, pH 7.2-7.4, 0.2 μm filtered. 10. Substrate Solution: Tetramethylbenzidine (Cat# AR1104) 11. Stop Solution: 2 N H2SO4 (Cat# AR1105) 12. Wash Buffer (PBS and PBS-T) . Recipes: PBS: 8g NaCl, 0.2g KCl, 1.15g Na2HPO4, 0.2g KH2PO4, adjust the total volume to 1 L with distilled water, pH 7.2-7.4, 0.2 μm filtered. PBS-T: 0.1% Tween® 20 in PBS, pH 7.2-7.4. *Item 6 – 12 are included in the EZ Set Accessory Kit (EZA001)