Mouse 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
    0.01 ng/mL
  • Assay Range
    0.1 - 5 ng/mL (Cell Culture Supernates, Serum, Platelet-poor EDTA Plasma, Platelet-poor Heparin Plasma)
  • Specificity
    Natural and recombinant mouse PF4
  • 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 PF4 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure mouse PF4 in cell culture supernates, serum, and platelet-poor plasma. It contains E. coli-expressed recombinant mouse PF4 and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant factor accurately. Results obtained using natural mouse PF4 showed dose-response curves that were parallel to the standard curves obtained using the recombinant kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse PF4.

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
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean0.230.411.510.250.451.46
Standard Deviation0.0130.0290.0460.0190.0350.11
CV%5.77.137.67.87.5

Recovery

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

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=4) 102 90-120
Linearity
To assess the linearity of the assay, samples containing high concentrations of mouse PF4 were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Mouse 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. 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.

    3 Citations: Showing 1 - 3
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    Species
    Sample Type
    1. Calf Spleen Extractive Injection protects mice against cyclophosphamide-induced hematopoietic injury through G-CSF-mediated JAK2/STAT3 signaling
      Authors: W Lu, D Jia, S An, M Mu, X Qiao, Y Liu, X Li, D Wang
      Sci Rep, 2017;7(1):8402.
      Species: Mouse
      Sample Type: Plasma
    2. Platelet Activation and Thrombus Formation over IgG Immune Complexes Requires Integrin alphaIIbbeta3 and Lyn Kinase.
      Authors: Zhi H, Dai J, Liu J, Zhu J, Newman D, Gao C, Newman P
      PLoS ONE, 2015;10(8):e0135738.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    3. Restoration of responsiveness of phospholipase Cgamma2-deficient platelets by enforced expression of phospholipase Cgamma1.
      Authors: Zheng Y, Adams T, Zhi H, Yu M, Wen R, Newman P, Wang D, Newman D
      PLoS ONE, 2015;10(3):e0119739.
      Species: Mouse
      Sample Type: Cell Culture Supernates
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