Human Proprotein Convertase 9/PCSK9 Quantikine ELISA Kit

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DPC900
PDPC900
SPC900
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Human Proprotein Convertase 9/PCSK9 ELISA Standard Curve
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Citations (24)
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Human Proprotein Convertase 9/PCSK9 Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Cell Lysates (50 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL)
Sensitivity
0.219 ng/mL
Assay Range
0.6 - 40 ng/mL (Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma)
Specificity
Natural and recombinant human PCSK9. This assay recognizes free and LDLR-bound PCSK9.
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.

Product Summary

The Quantikine Human PCSK9 immunoassay is a 4.5 hour solid phase ELISA designed to measure PCSK9 in cell culture supernates, cell lysates, serum, and plasma. It contains NS0-expressed recombinant human PCSK9 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural PCSK9 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring PCSK9.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision

Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean 4.82 14 27.7 4.64 14.5 27.9
Standard Deviation 0.196 0.795 1.81 0.276 0.629 1.13
CV% 4.1 5.7 6.5 6 4.3 4.1

Recovery

The recovery of PCSK9 spiked to levels throughout the range of the assay was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 107 100-111

Linearity

To assess the linearity of the assay, samples containing high concentrations of PCSK9 were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human Proprotein Convertase 9/PCSK9 ELISA Linearity

Data Examples

Human Proprotein Convertase 9/PCSK9 ELISA Standard Curve

Product Datasheets

Preparation and Storage

Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: Proprotein Convertase 9/PCSK9

Proprotein convertase subtilisin kexin 9 (PCSK9), also named neural apoptosis-regulated convertase 1 (NARC-1), is a member of the proteinase K subfamily of subtilisin-related serine endoproteases. The full-length protein has 692 amino acids, including a signal peptide, a pro- domain, and a catalytic domain. PCSK9 is highly expressed in the liver, intestine, and kidney. It is initially synthesized as a soluble 74 kDa precursor protein. In the endoplasmic reticulum, it undergoes autocatalytic intramolecular cleavage to generate a 14 kDa pro- domain and a 60 kDa catalytic domain. These two domains remain associated when PCSK9 is secreted outside the cells (1-3). The primary physiologic function of PCSK9 is to mediate the degradation of low density lipoprotein receptor (LDL R). Early observations indicated that gain-of-function missense mutations in the PCSK9 gene can cause an autosomal dominant form of hypercholesterolemia (4, 5). The expression of PCSK9 was observed to be up-regulated by the sterol regulatory element binding proteins (SREBPs), a family of transcription factors that are responsible for the upregulation of genes involved in cholesterol and fatty acid metabolism, such as the LDL R gene (6, 7). Further experimental evidence revealed that in mice, when the PCSK9 gene was knocked out, the number of LDL R in hepatocytes increased, whereas when PCSK9 was over-expressed, the amount of LDL R protein was reduced in the liver (8, 9). In humans, genetic analyses have shown that individuals who have nonsense or loss-of-function mutations in the PCSK9 gene have significantly lower plasma LDL cholesterol levels (10, 11). These investigations clearly indicated that PCSK9 plays a key role in reducing the hepatic LDL R levels. Recently, the underlying mechanism has been uncovered: under normal physiologic conditions, the LDL R is internalized on the cell surface and directed to the endosomes in order to be recycled back to the cell surface. PCSK9 binds to the EGF domain of the LDL R and prevents LDL R from being sorted to the endosomes. Instead, the PCSK9/LDL R complex is redistributed to the lysosomes for degradation (12-14). As such, PCSK9 regulates the amount of LDL R in the circulation and modulates cholesterol levels. Serum PCSK9 concentrations have been found to be directly associated with cholesterol levels (15, 16). Since individuals with loss-of-function PCSK9 mutations have strikingly reduced risk of coronary heart diseases, PCSK9 has become an attractive drug target in recent years (17, 18). One approach is to generate small molecules that are able to interfere with PCSK9 autoactivation and its interaction with LDL R. Other approaches aiming to reduce the amounts of PCSK9 in the circulation, such as small interfering RNAs (siRNAs), have also shown promise (19, 20).

Entrez Gene IDs:
255738 (Human); 100102 (Mouse); 298296 (Rat); 102142788 (Cynomolgus Monkey)
Alternate Names:
EC 3.4.21; EC 3.4.21.111; FH3; FH3neural apoptosis regulated convertase 1; FHCL3; HCHOLA3; hypercholesterolemia, autosomal dominant 3; LDLCQ1; NARC1; NARC-1; NARC-1convertase subtilisin/kexin type 9 preproprotein; NARC1EC 3.4.21.-; Neural apoptosis-regulated convertase 1; PC9; PCSK9; Proprotein Convertase 9; proprotein convertase subtilisin/kexin type 9; Subtilisin/kexin-like protease PC9
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Assay Procedure

Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 100 µL Assay Diluent
  4.   Add 100 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 200 µL Conjugate
  9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  10.   Aspirate and wash 4 times.

  11. 200 µL Substrate Solution
  12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

  13. 50 µL Stop Solution
  14.   Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

Citations for Human Proprotein Convertase 9/PCSK9 Quantikine ELISA Kit

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

24 Citations: Showing 1 - 10
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  1. Familial hypercholesterolemia: Is there a role for PCSK9 and thrombin generation?
    Authors: JPP Silvino, MG Carvalho, EA Reis, APL Mota, KB Gomes, RCF Duarte, MCJ Guimarães, MCR Sousa, PS Azevedo, IFO Silva
    Thrombosis Research, 2021;200(0):156-163.
    Species: Human
    Sample Types: Plasma
  2. PCSK9 Levels Are Raised in Chronic HCV Patients with Hepatocellular Carcinoma
    Authors: S Fasolato, S Pigozzo, P Pontisso, P Angeli, M Ruscica, E Savarino, S De Martin, MG Lupo, N Ferri
    J Clin Med, 2020;9(10):.
    Species: Human
    Sample Types: Plasma
  3. Methotrexate Decreases the Level of PCSK9-A Novel Indicator of the Risk of Proatherogenic Lipid Profile in Psoriasis. The Preliminary Data
    Authors: JA Krahel, A Baran, TW Kami?ski, M Maciaszek, I Flisiak
    J Clin Med, 2020;9(4):.
    Species: Human
    Sample Types: Whole Blood
  4. Beneficial impact of epigallocatechingallate on LDL-C through PCSK9/LDLR pathway by blocking HNF1&alpha and activating FoxO3a
    Authors: CJ Cui, JL Jin, LN Guo, J Sun, NQ Wu, YL Guo, G Liu, Q Dong, JJ Li
    J Transl Med, 2020;18(1):195.
    Species: Human
    Sample Types: Plasma
  5. Circulating PCSK9 and cardiovascular events in FH patients with standard lipid-lowering therapy
    Authors: YX Cao, JL Jin, D Sun, HH Liu, YL Guo, NQ Wu, RX Xu, CG Zhu, Q Dong, J Sun, JJ Li
    J Transl Med, 2019;17(1):367.
    Species: Human
    Sample Types: Plasma
  6. Regulation of Cholesterol Homeostasis by a Novel Long Non-coding RNA LASER
    Authors: C Li, Z Hu, W Zhang, J Yu, Y Yang, Z Xu, H Luo, X Liu, Y Liu, C Chen, Y Cai, X Xia, X Zhang, DZ Wang, G Wu, C Zeng
    Sci Rep, 2019;9(1):7693.
    Species: Human
    Sample Types: Cell Lysates
  7. In vivo genome and base editing of a human PCSK9 knock-in hypercholesterolemic mouse model
    Authors: A Carreras, LS Pane, R Nitsch, K Madeyski-B, M Porritt, P Akcakaya, A Taheri-Gha, E Ericson, M Bjursell, M Perez-Alca, F Seeliger, M Althage, R Knöll, R Hicks, LM Mayr, R Perkins, D Lindén, J Borén, M Bohlooly-Y, M Maresca
    BMC Biol., 2019;17(1):4.
    Species: Transgenic Mouse
    Sample Types: Plasma
  8. In vivo CRISPR editing with no detectable genome-wide off-target mutations
    Authors: P Akcakaya, ML Bobbin, JA Guo, J Malagon-Lo, K Clement, SP Garcia, MD Fellows, MJ Porritt, MA Firth, A Carreras, T Baccega, F Seeliger, M Bjursell, SQ Tsai, NT Nguyen, R Nitsch, LM Mayr, L Pinello, M Bohlooly-Y, MJ Aryee, M Maresca, JK Joung
    Nature, 2018;0(0):.
    Species: Mouse
    Sample Types: Plasma
  9. FXR Activation by Obeticholic Acid or Non-Steroidal Agonists Induces a Human-Like Lipoprotein Cholesterol Change in Mice with Humanized Chimeric Liver
    Authors: R Papazyan, X Liu, J Liu, B Dong, EM Plummer, RD Lewis, JD Roth, MA Young
    J. Lipid Res., 2018;0(0):.
    Species: Mouse
    Sample Types: Serum
  10. Impact of PCSK9 loss-of-function genotype on 1-year mortality and recurrent infection in sepsis survivors
    Authors: KR Genga, C Lo, MS Cirstea, FS Leitao Fil, KR Walley, JA Russell, A Linder, GA Francis, JH Boyd
    EBioMedicine, 2018;0(0):.
    Species: Human
    Sample Types: Plasma
  11. PCSK9 induces a pro-inflammatory response in macrophages
    Authors: C Ricci, M Ruscica, M Camera, L Rossetti, C Macchi, A Colciago, I Zanotti, MG Lupo, MP Adorni, AFG Cicero, F Fogacci, A Corsini, N Ferri
    Sci Rep, 2018;8(1):2267.
    Species: Human
    Sample Types: Cell Culture Supernates
  12. Loss-of-function PCSK9 mutants evade the unfolded protein response sensor, GRP78, and fail to induce endoplasmic reticulum stress when retained
    Authors: P Lebeau, K Platko, AA Al-Hashimi, JH Byun, Š Lhoták, N Holzapfel, G Gyulay, SA Igdoura, D Cool, B Trigatti, NG Seidah, RC Austin
    J. Biol. Chem., 2018;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  13. Relationship of PCSK9 levels with indices of vascular function and subclinical atherosclerosis in patients with familial dyslipidaemias
    Authors: C Vlachopoul, I Koutagiar, D Terentes-P, I Skoumas, A Rigatou, A Miliou, AN Skliros, S Pantou, K Filis, D Tousoulis
    Hellenic J Cardiol, 2018;0(0):.
    Species: Human
    Sample Types: Plasma
  14. Ossabaw Pigs With a PCSK9 Gain-of-Function Mutation Develop Accelerated Coronary Atherosclerotic Lesions: A Novel Model for Preclinical Studies
    Authors: F Yuan, L Guo, KH Park, JR Woollard, K Taek-Geun, K Jiang, T Melkamu, B Zang, SL Smith, SC Fahrenkrug, FD Kolodgie, A Lerman, R Virmani, LO Lerman, DF Carlson
    J Am Heart Assoc, 2018;7(6):.
    Species: Porcine
    Sample Types: Plasma
  15. Heparan sulfate proteoglycans present PCSK9 to the LDL receptor
    Authors: C Gustafsen, D Olsen, J Vilstrup, S Lund, A Reinhardt, N Wellner, T Larsen, CBF Andersen, K Weyer, JP Li, PH Seeberger, S Thirup, P Madsen, S Glerup
    Nat Commun, 2017;8(1):503.
    Species: Human
    Sample Types: Cell Culture Supernates
  16. Circulating Levels of Proprotein Convertase Subtilisin/Kexin Type 9 and Arterial Stiffness in a Large Population Sample: Data From the Brisighella Heart Study
    Authors: M Ruscica, N Ferri, F Fogacci, M Rosticci, M Botta, S Marchiano, P Magni, S D'Addato, M Giovannini, C Borghi, AFG Cicero,
    J Am Heart Assoc, 2017;6(5):.
    Species: Human
    Sample Types: Serum
  17. CAT-2003: A novel sterol regulatory element-binding protein inhibitor that reduces steatohepatitis, plasma lipids, and atherosclerosis in apolipoprotein E*3-Leiden mice
    Authors: M Zimmer, P Bista, EL Benson, DY Lee, F Liu, D Picarella, RB Vega, CB Vu, M Yeager, M Ding, G Liang, JD Horton, R Kleemann, T Kooistra, MC Morrison, PY Wielinga, JC Milne, MR Jirousek, AJ Nichols
    Hepatol Commun, 2017;1(4):311-325.
    Species: Human
    Sample Types: Cell Culture Supernates
  18. Proprotein Convertase Subtilisin/Kexin 9 Levels in Relation to Systemic Immune Activation and Subclinical Coronary Plaque in HIV
    Authors: MV Zanni, LA Stone, M Toribio, DE Rimmelin, J Robinson, TH Burdo, K Williams, KV Fitch, J Lo, SK Grinspoon
    Open Forum Infect Dis, 2017;4(4):ofx227.
    Species: Human
    Sample Types: Serum
  19. Endoplasmic reticulum stress and Ca2+ depletion differentially modulate the sterol-regulatory protein PCSK9 to control lipid metabolism
    Authors: Paul Lebeau
    J. Biol. Chem, 2016;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  20. A MARCH6 and IDOL E3 Ubiquitin Ligase Circuit Uncouples Cholesterol Synthesis from Lipoprotein Uptake in Hepatocytes.
    Authors: Loregger A, Cook E, Nelson J, Moeton M, Sharpe L, Engberg S, Karimova M, Lambert G, Brown A, Zelcer N
    Mol Cell Biol, 2016;36(2):285-94.
    Species: Human
    Sample Types: Cell Culture Supernates
  21. Plasma PCSK9 level is unrelated to blood pressure and not associated independently with carotid intima-media thickness in hypertensives
    Authors: SH Yang, Y Du, S Li, Y Zhang, RX Xu, CG Zhu, YL Guo, NQ Wu, Q Dong, J Sun, JJ Li
    Hypertens Res, 2016;0(0):.
    Species: Human
    Sample Types: Plasma
  22. Plasma PCSK9 levels are elevated with acute myocardial infarction in two independent retrospective angiographic studies.
    Authors: Almontashiri N, Vilmundarson R, Ghasemzadeh N, Dandona S, Roberts R, Quyyumi A, Chen H, Stewart A
    PLoS ONE, 2014;9(9):e106294.
    Species: Human
    Sample Types: Plasma
  23. Impact of currently prescribed lipid-lowering drugs on plasma PCSK9 concentration: single or in combination study in rats.
    Authors: Zhang Y, Liu J, Li S, Xu R, Sun J, Li J
    Lipids Health Dis, 2014;13(0):35.
    Species: Rat
    Sample Types: Plasma
  24. Circulating Levels of Proprotein Convertase Subtilisin/Kexin Type 9 and Arterial Stiffness in a Large Population Sample: Data From the Brisighella Heart Study.
    Authors: Ruscica M, Ferri N, Fogacci F, Rosticci M, Botta M, Marchiano S, Magni P, D'Addato S, Giovannini M, Borghi C, Cicero A
    J Am Heart Assoc, 0;6(5):.
    Species: Human
    Sample Types: Plasma

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Human Proprotein Convertase 9/PCSK9 Quantikine ELISA Kit
By Anonymous on 05/28/2018
Application: Sample Tested: Cell Culture Media