Human Proprotein Convertase 9/PCSK9 Quantikine ELISA Kit

  (12 citations)     
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Product Details
Assay Procedure
Citations (12)
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  • 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.
Control Available
QC235 , Quantikine Immunoassay Control Group 235 - Please Inquire
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.
 Proprotein Convertase 9/PCSK9 [HRP]
Product Datasheets

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Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: Proprotein Convertase 9/PCSK9
This family of serine proteases consists of furin and proprotein convertases encoded by PCSK genes. They are important in a variety of processes. Furin has an essential role in embryogenesis and homeostasis and is implicated in various pathologies such as cancer, neurodegenerative diseases and anthrax. Mutations in the PC1/PCSK1 gene are associated with obesity and impaired prohormone processing.
  • Entrez Gene IDs:
    255738 (Human); 100102 (Mouse); 298296 (Rat)
  • Alternate Names:
    EC 3.4.21; EC 3.4.21.111; FH3; FH3neural apoptosis regulated convertase 1; HCHOLA3; hypercholesterolemia, autosomal dominant 3; LDLCQ1; 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
Related Research Areas
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:

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.

12 Citations: Showing 1 - 10
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Species
Sample Type
  1. 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 Type: Cell Culture Supernates
  2. 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 Type: Plasma
  3. 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 Type: Cell Culture Supernates
  4. 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 Type: Cell Culture Supernates
  5. 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 Type: Serum
  6. 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 Type: Serum
  7. 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 Type: Cell Culture Supernates
  8. 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 Type: Plasma
  9. 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 Type: Cell Culture Supernates
  10. 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 Type: Plasma
  11. 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 Type: Plasma
  12. 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 Type: Plasma
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ELISA Controls
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Quantikine Immunoassay Control Group 235

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Quantikine ELISA Wash Buffer 1

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Cell Lysis Buffer 1 (1 x 21 mL)

ELISA 890713 1
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