Human CXCL4/PF4 DuoSet ELISA

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Human CXCL4 / PF4 ELISA Standard Curve
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Product Details
Citations (23)
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Human CXCL4/PF4 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
96-well strip plate
Sample Volume Required
100 µL
Assay Range
15.6 - 1,000 pg/mL
Sufficient Materials
For fifteen 96-well plates*
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human CXCL4/PF4. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Scientific Data

Human CXCL4 / PF4 ELISA Standard Curve

Product Datasheets

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Preparation and Storage

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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

Assay Procedure


Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

Citations for Human CXCL4/PF4 DuoSet ELISA

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.

23 Citations: Showing 1 - 10
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  1. In Vitro Comparison of Lymphangiogenic Potential of Hypoxia Preconditioned Serum (HPS) and Platelet-Rich Plasma (PRP)
    Authors: J Jiang, X Cong, S Alageel, U Dornseifer, AF Schilling, E Hadjipanay, HG Machens, P Moog
    International Journal of Molecular Sciences, 2023-01-19;24(3):.
    Species: Human
    Sample Types: Hypoxia Preconditioned Serum
  2. CXCL4-RNA Complexes Circulate in Systemic Sclerosis and Amplify Inflammatory/Pro-Fibrotic Responses by Myeloid Dendritic Cells
    Authors: I Pietrafort, A Butera, L Gaddini, A Mennella, R Palazzo, D Campanile, K Stefananto, V Riccieri, R Lande, L Frasca
    International Journal of Molecular Sciences, 2022-12-30;24(1):.
    Species: Human
    Sample Types: Plasma
  3. The effect of transdermal gender-affirming hormone therapy on markers of inflammation and hemostasis
    Authors: MH Schutte, R Kleemann, NM Nota, CM Wiepjes, JM Snabel, G T'Sjoen, A Thijs, M den Heijer
    PLoS ONE, 2022-03-15;17(3):e0261312.
    Species: Human
    Sample Types: Serum
  4. Intramammary treatment using allogeneic pure platelet-rich plasma in cows with subclinical mastitis caused by Gram-positive bacteria
    Authors: PC Duque-Madr, J Velasco-Bo, A Ceballos-M, C López, JU Carmona
    Scientific Reports, 2021-12-09;11(1):23737.
    Species: Bovine
    Sample Types: Plasma
  5. Rapid Internalization and Nuclear Translocation of CCL5 and CXCL4 in Endothelial Cells
    Authors: A Dickhout, DM Kaczor, ACA Heinzmann, SLN Brouns, JWM Heemskerk, MAMJ van Zandvo, RR Koenen
    International Journal of Molecular Sciences, 2021-07-08;22(14):.
    Species: Human
    Sample Types: Cell Lysates
  6. Galectin-1 and platelet factor 4 (CXCL4) induce complementary platelet responses in vitro
    Authors: A Dickhout, BME Tullemans, JWM Heemskerk, VLJL Thijssen, MJE Kuijpers, RR Koenen
    PLoS ONE, 2021-01-07;16(1):e0244736.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. Multiple Tissue Biomarkers Independently and Additively Predict Prostate Cancer Pathology Outcomes
    Authors: MR Cooperberg, JE Cowan, KJ Lindquist, Y Kobayashi, JP Simko, H Bengtsson, K Singh, V Ngo, A Avila, LF Newcomb, M Tretriakov, DW Lin, S Stone, PR Carroll, PL Paris
    Eur Urol, 2020-11-02;0(0):.
    Species: Human
    Sample Types: Plasma
  8. CXCL4 assembles DNA into liquid crystalline complexes to amplify TLR9-mediated interferon-? production in systemic sclerosis
    Authors: R Lande, EY Lee, R Palazzo, B Marinari, I Pietrafort, GS Santos, Y Mattenberg, F Spadaro, K Stefananto, N Iannace, AM Dufour, M Falchi, M Bianco, E Botti, L Bianchi, M Alvarez, V Riccieri, ME Truchetet, G C L Wong, C Chizzolini, L Frasca
    Nat Commun, 2019-05-01;10(1):1731.
    Species: Human
    Sample Types: Plasma
  9. Optimizing Measurement of Vascular Endothelial Growth Factor in Small Blood Samples of Premature Infants
    Authors: CC Lopez Yoma, KT Preissner, B Lorenz, K Stieger
    Sci Rep, 2019-05-01;9(1):6744.
    Species: Human
    Sample Types: Serum
  10. Characterisation of the Ral GTPase inhibitor RBC8 in human and mouse platelets
    Authors: TG Walsh, A Wersäll, AW Poole
    Cell. Signal., 2019-03-14;0(0):.
    Species: Human
    Sample Types: Cell Lysates
  11. Inhibition of profibrotic microRNA-21 affects platelets and their releasate
    Authors: T Barwari, S Eminaga, U Mayr, R Lu, PC Armstrong, MV Chan, M Sahraei, M Fernández-, T Moreau, J Barallobre, M Lynch, X Yin, C Schulte, F Baig, R Pechlaner, SR Langley, A Zampetaki, P Santer, M Weger, R Plasenzott, M Schosserer, J Grillari, S Kiechl, J Willeit, AM Shah, C Ghevaert, TD Warner, C Fernández-, Y Suárez, M Mayr
    JCI Insight, 2018-11-02;3(21):.
    Species: Human
    Sample Types: Plasma
  12. Preemptively and non-preemptively transplanted patients show a comparable hypercoagulable state prior to kidney transplantation compared to living kidney donors
    Authors: GJ Nieuwenhui, TAJ van den Be, SJL Bakker, MC van den He, MMRF Struys, T Lisman, RA Pol
    PLoS ONE, 2018-07-16;13(7):e0200537.
    Species: Human
    Sample Types: Plasma
  13. Urinary angiostatin, CXCL4 and VCAM-1 as biomarkers of lupus nephritis
    Authors: CC Mok, S Soliman, LY Ho, FA Mohamed, FI Mohamed, C Mohan
    Arthritis Res. Ther., 2018-01-11;20(1):6.
    Species: Human
    Sample Types: Urine
  14. A combination of platelet features allows detection of early-stage cancer
    Authors: S Sabrkhany, MJE Kuijpers, SMJ van Kuijk, L Sanders, S Pineda, SWM Olde Damin, AC Dingemans, AW Griffioen, MGA Oude Egbri
    Eur. J. Cancer, 2017-05-17;80(0):5-13.
    Species: Human
    Sample Types: Platelets
  15. Vascular endothelial growth factor A is associated with the subsequent development of moderate or severe cardiac allograft vasculopathy in pediatric heart transplant recipients
    Authors: David M Briscoe
    J. Heart Lung Transplant., 2016-10-03;0(0):.
    Species: Human
    Sample Types: Plasma
  16. Novel Antiplatelet Activity of Minocycline Involves Inhibition of MLK3-p38 Mitogen Activated Protein Kinase Axis
    Authors: Joseph W Jackson
    PLoS ONE, 2016-06-06;11(6):e0157115.
    Species: Human
    Sample Types: Plasma
  17. Host biomarkers distinguish dengue from leptospirosis in Colombia: a case-control study.
    Authors: Conroy A, Gelvez M, Hawkes M, Rajwans N, Liles W, Villar-Centeno L, Kain K
    BMC Infect Dis, 2014-01-20;14(0):35.
    Species: Human
    Sample Types: Serum
  18. Proteome-wide analysis and CXCL4 as a biomarker in systemic sclerosis
    Authors: L van Bon, AJ Affandi, J Broen, RB Christmann, RJ Marijnisse, L Stawski, GA Farina, G Stifano, AL Mathes, M Cossu, M York, C Collins, M Wenink, R Huijbens, R Hesselstra, T Saxne, M DiMarzio, D Wuttge, SK Agarwal, JD Reveille, S Assassi, M Mayes, Y Deng, JP Drenth, J de Graaf, M den Heijer, CG Kallenberg, M Bijl, A Loof, WB van den Be, LA Joosten, V Smith, F de Keyser, R Scorza, C Lunardi, PL van Riel, M Vonk, W van Heerde, S Meller, B Homey, L Beretta, M Roest, M Trojanowsk, R Lafyatis, TR Radstake
    N. Engl. J. Med, 2013-12-18;370(5):433-43.
    Species: Human
    Sample Types: Plasma
  19. Distinct platelet packaging, release, and surface expression of proangiogenic and antiangiogenic factors on different platelet stimuli.
    Authors: Chatterjee M, Huang Z, Zhang W, Jiang L, Hultenby K, Zhu L, Hu H, Nilsson GP, Li N
    Blood, 2011-02-17;117(14):3907-11.
    Species: Human
    Sample Types: Cell Culture Supernates
  20. Thrombocytopenia in early malaria is associated with GP1b shedding in absence of systemic platelet activation and consumptive coagulopathy.
    Authors: de Mast Q, de Groot PG, van Heerde WL, Roestenberg M, van Velzen JF, Verbruggen B, Roest M, McCall M, Nieman AE, Westendorp J, Syafruddin D, Fijnheer R, van Dongen-Lases EC, Sauerwein RW, van der Ven AJ
    Br. J. Haematol., 2010-10-19;151(5):495-503.
    Species: Human
    Sample Types: Plasma
  21. Functional divergence between 2 chemokines is conferred by single amino acid change.
    Authors: Dubrac A, Quemener C, Lacazette E, Lopez F, Zanibellato C, Wu WG, Bikfalvi A, Prats H
    Blood, 2010-08-05;116(22):4703-11.
    Species: Human
    Sample Types: Cell Culture Supernates
  22. Normal ranges of angiogenesis regulatory proteins in human platelets.
    Authors: Peterson JE, Zurakowski D, Italiano JE
    Am. J. Hematol., 2010-07-01;85(7):487-93.
    Species: Human
    Sample Types: Cell Lysates
  23. BMP4 regulation of human megakaryocytic differentiation is involved in thrombopoietin signaling.
    Authors: Jeanpierre S, Nicolini FE, Kaniewski B, Dumontet C, Rimokh R, Puisieux A, Maguer-Satta V
    Blood, 2008-07-29;112(8):3154-63.
    Species: Human
    Sample Types: Cell Culture Supernates


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Human CXCL4/PF4 DuoSet ELISA
By Anonymous on 10/02/2018
Sample Tested: Serum and Plasma