Human CXCL4/PF4 Quantikine ELISA Kit

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DPF40
Control Products Available
Human CXCL4/PF4 ELISA Standard Curve
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Procedure
Citations (12)
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Human CXCL4/PF4 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), 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.

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
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (ng/mL) 5.8 14.1 26.8 5.61 14.3 27.2
Standard Deviation 0.472 0.84 1.85 0.558 1.7 3.2
CV% 8.1 6 6.9 9.9 11.9 11.8

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 ELISA Linearity

Scientific Data

Human CXCL4/PF4 ELISA Standard Curve

Product Datasheets

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Preparation and Storage

Shipping
The product is shipped at ambient temperature. 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: 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
⚠ 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 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 for Human CXCL4/PF4 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.

12 Citations: Showing 1 - 10
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  1. Heparin-Functionalized Adsorbents Eliminate Central Effectors of Immunothrombosis, including Platelet Factor 4, High-Mobility Group Box 1 Protein and Histones
    Authors: M Ebeyer-Mas, T Eichhorn, R Weiss, V Semak, L Lauková, MB Fischer, V Weber
    International Journal of Molecular Sciences, 2022;23(3):.
    Species: Human
    Sample Types: Plasma
  2. Immune responses in Omicron SARS-CoV-2 breakthrough infection in vaccinated adults
    Authors: H Kared, AS Wolf, A Alirezayla, A Ravussin, G Solum, TT Tran, F Lund-Johan, JT Vaage, LS Nissen-Mey, UC Nygaard, O Hungnes, AH Robertson, LM Næss, L Trogstad, P Magnus, LA Munthe, S Mjaaland
    Nature Communications, 2022;13(1):4165.
    Species: Human
    Sample Types: Plasma
  3. Differences in the Biomarker Profile of De Novo Acute Heart Failure versus Decompensation of Chronic Heart Failure
    Authors: S Nawrocka-M, J Biegus, M Hurkacz, M Guzik, M Rosiek-Bie, EA Jankowska, P Ponikowski, R Zymli?ski
    Biomolecules, 2021;11(11):.
    Species: Human
    Sample Types: Plasma
  4. Elevated Lactate Levels in Acute Pulmonary Embolism Are Associated with Prothrombotic Fibrin Clot Properties: Contribution of NETs Formation
    Authors: M Z?bczyk, J Natorska, A Janion-Sad, KP Malinowski, M Janion, A Undas
    J Clin Med, 2020;9(4):.
    Species: Human
    Sample Types: Plasma
  5. Impact of Anticoagulation and Sample Processing on the Quantification of Human Blood-Derived microRNA Signatures
    Authors: M Mussbacher, TL Krammer, S Heber, WC Schrottmai, S Zeibig, HP Holthoff, D Pereyra, P Starlinger, M Hackl, A Assinger
    Cells, 2020;9(8):.
    Species: Human
    Sample Types: Plasma
  6. Impact of Anticoagulation and Sample Processing on the Quantification of Human Blood-Derived microRNA Signatures
    Authors: M Mussbacher, TL Krammer, S Heber, WC Schrottmai, S Zeibig, HP Holthoff, D Pereyra, P Starlinger, M Hackl, A Assinger
    Cells, 2020;9(8):.
    Species: Human
    Sample Types: Plasma
  7. Increased platelet factor 4 and aberrant permeability of follicular fluid in PCOS
    Authors: CC Huang, CH Chou, SU Chen, HN Ho, YS Yang, MJ Chen
    J. Formos. Med. Assoc., 2018;0(0):.
    Species: Human
    Sample Types: Follicular Fluid
  8. Optimized plasma preparation is essential to monitor platelet-stored molecules in humans
    Authors: M Mussbacher, WC Schrottmai, M Salzmann, C Brostjan, JA Schmid, P Starlinger, A Assinger
    PLoS ONE, 2017;12(12):e0188921.
    Species: Human
    Sample Types: Plasma
  9. Changes in plasma CXCL4 levels are associated with improvements in lung function in patients receiving immunosuppressive therapy for systemic sclerosis-related interstitial lung disease
    Authors: Elizabeth R Volkmann
    Arthritis Res. Ther, 2016;18(1):305.
    Species: Human
    Sample Types: Plasma
  10. 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 Types: Plasma
  11. 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 Types: Cell Culture Supernates
  12. 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 Types: Cell Culture Supernates

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Reviews for Human CXCL4/PF4 Quantikine ELISA Kit

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Human CXCL4/PF4 Quantikine ELISA Kit
By Anonymous on 09/14/2019
Sample Tested: Recombinant protein

Human CXCL4/PF4 Quantikine ELISA Kit
By Balaji Mahender on 01/09/2018
Sample Tested: Citrate Plasma,EDTA Plasma