Mouse Proprotein Convertase 9/PCSK9 Quantikine ELISA Kit

  (6 citations)
(1 Review)
    
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Product Details
Assay Procedure
Citations (6)
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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 (10 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL)
  • Sensitivity
    21.9 pg/mL
  • Assay Range
    62.5 - 4,000 pg/mL (Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma)
  • Specificity
    Natural and recombinant mouse PCSK9. This kit detects 60 kDa, 53 kDa, and LDLR-complexed recombinant mouse 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 Mouse PCSK9 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure mouse PCSK9 in cell culture supernates, cell lysates, serum, and plasma. It contains NS0-expressed recombinant mouse PCSK9 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural mouse 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 mouse 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 181 427 1427 213 469 1511
Standard Deviation 10.3 32.1 44.1 18.9 25.9 102
CV% 5.7 7.5 3.1 8.9 5.5 6.8

Recovery

The recovery of mouse PCSK9 spiked to three levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=6) 102 94-115
Cell Lysates (n=5) 103 91-117
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of mouse PCSK9 in each matrix were diluted with Calibrator Diluent and then assayed.
 Proprotein Convertase 9/PCSK9 [HRP]
Product Datasheets

Certificate of Analysis Lookup

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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. 50 µL Assay Diluent
  4.   Add 50 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 4 times for a total of 5 washes.

  8. 100 µL Conjugate
  9.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  10.   Aspirate and wash 5 times.

  11. 100 µL Substrate Solution
  12.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

  13. 100 µL Stop Solution
  14.   Add 100 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.

6 Citations: Showing 1 - 6
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Species
Sample Type
  1. Development of vaccine for dyslipidemia targeted to a proprotein convertase subtilisin/kexin type 9 (PCSK9) epitope in mice
    Authors: R Kawakami, Y Nozato, H Nakagami, Y Ikeda, M Shimamura, S Yoshida, J Sun, T Kawano, Y Takami, T Noma, H Rakugi, T Minamino, R Morishita
    PLoS ONE, 2018;13(2):e0191895.
    Species: Mouse
    Sample Type: Plasma
  2. 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: Mouse
    Sample Type: Plasma
  3. Regulation of lipid metabolism by obeticholic acid in hyperlipidemic hamsters
    Authors: Bin Dong
    J. Lipid Res, 2016;0(0):.
    Species: Hamster
    Sample Type: Serum
  4. Inhibition of PCSK9 transcription by berberine involves down-regulation of hepatic HNF1alpha protein expression through the ubiquitin-proteasome degradation pathway.
    Authors: Dong B, Li H, Singh A, Cao A, Liu J
    J Biol Chem, 2015;290(7):4047-58.
    Species: Hamster
    Sample Type: Serum
  5. Reduction of circulating PCSK9 and LDL-C levels by liver-specific knockdown of HNF1alpha in normolipidemic mice.
    Authors: Shende V, Wu M, Singh A, Dong B, Kan C, Liu J
    J Lipid Res, 2015;56(4):801-9.
    Species: Mouse
    Sample Type: Serum
  6. Permanent alteration of PCSK9 with in vivo CRISPR-Cas9 genome editing.
    Authors: Ding Q, Strong A, Patel K, Ng S, Gosis B, Regan S, Cowan C, Rader D, Musunuru K
    Circ Res, 2014;115(5):488-92.
    Species: Mouse
    Sample Type: Plasma

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