Human CXCL4/PF4 Quantikine ELISA Kit

(3 citations)   
  • 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), Serum (10 uL), Platelet-poor EDTA Plasma (10 uL), Platelet-poor Heparin Plasma (10 uL)
  • Sensitivity
    0.1 ng/mL
  • Assay Range
    0.8 - 50 ng/mL (Cell Culture Supernates, Serum, Platelet-poor EDTA Plasma, Platelet-poor Heparin Plasma)
  • Specificity
    Natural and recombinant human PF4.
  • Cross-reactivity
    Cross-reactivity observed with 1 or more available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    No significant interference observed with available related molecules.
Control Available
QC141, Quantikine Immunoassay Control Set 800 for Human CXCL4/PF4 - Please Inquire
Product Summary
The Quantikine Human CXCL4/PF4 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human PF4 in cell culture supernates, serum, and platelet-poor plasma. It contains E. coli-expressed recombinant human PF4 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human PF4 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 human PF4.

Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components.
Cell Culture Supernates, Serum, Platelet-poor EDTA Plasma, Platelet-poor Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean5.814.126.85.6114.327.2
Standard Deviation0.4720.841.850.5581.73.2
CV%8.166.99.911.911.8

Recovery

The recovery of human PF4 spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 110 99-117
Platelet-poor EDTA Plasma (n=4) 100 89-117
Linearity
To assess linearity of the assay, samples containing and/or spiked with high concentrations of human PF4 were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human CXCL4/PF4 Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: CXCL4/PF4
CXCL4/PF4 is a secreted chemokine that is expressed by megakaryocytes and restrains the coagulation cascade. It is released by activated platelets and regulates Thrombin/Thrombomodulin complexes, activated Protein C (APC), Coagulation Factor Xa, and Fibrin. The anticoagulant heparin neutralizes CXCL4 procoagulant effects. Complexes of heparin and CXCL4 trigger antibody formation that causes the pathological syndrome HITT. Immunogenic complexes of CXCL4 with Apolipoprotein H contribute to antiphospholipid syndrome (APS). CXCL4 interferes with the proliferative and angiogenic functions of FGF-2 and VEGF. However, it can also exert proinflammatory and pro-atherogenic effects on monocytes, macrophages, and endothelial cells. CXCL4 may signal through CXCR3B in humans.
    • Entrez Gene IDs
      5196 (Human); 56744 (Mouse); 360918 (Rat);
    • Alternate Names
      chemokine (C-X-C motif) ligand 4; C-X-C motif chemokine 4; CXCL4; CXCL4iroplact; Iroplact; MGC138298; Oncostatin-A; PF4; PF-4; platelet factor 4; SCYB4oncostatin-A;
    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 on a horizontal orbital microplate shaker.
    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 on the shaker.
    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 on the benchtop. 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.

    3 Citations: Showing 1 - 3
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    Species
    Sample Type
    1. Unaltered Angiogenesis-Regulating Activities of Platelets in Mild Type 2 Diabetes Mellitus despite a Marked Platelet Hyperreactivity
      PLoS ONE, 2016;11(9):e0162405.
      Species: Human
      Sample Type: Cell Culture Supernates
    2. Smoothened Agonist Reduces Human Immunodeficiency Virus Type-1-Induced Blood-Brain Barrier Breakdown in Humanized Mice
      Authors: Vir B Singh
      Sci Rep, 2016;6(0):26876.
      Species: Mouse
      Sample Type: Plasma
    3. Endothelial activation by platelets from sickle cell anemia patients.
      Authors: Proenca-Ferreira R, Brugnerotto A, Garrido V, Dominical V, Vital D, Ribeiro M, dos Santos M, Traina F, Olalla-Saad S, Costa F, Conran N
      PLoS ONE, 2014;9(2):e89012.
      Species: Human
      Sample Type: Cell Culture Supernates
    ELISA Controls
    Description Application Cat# Citations Images  

    Quantikine Immunoassay Control Set 800 for Human CXCL4/PF4

    Ctrl QC141
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    Supplemental ELISA Products
    Description Application Cat# Citations Images  

    Quantikine ELISA Wash Buffer 1

    ELISA WA126 6
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