Human CXCL4/PF4 DuoSet ELISA

Catalog # Availability Size / Price Qty
DY795
Ancillary Products Available
Human CXCL4 / PF4 ELISA Standard Curve
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Product Details
Procedure
Citations (11)
FAQs
Supplemental Products
Reviews (1)

Human CXCL4/PF4 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Sufficient Materials
For fifteen 96-well plates*
Specificity
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)

Data Example

Human CXCL4 / PF4 ELISA Standard Curve

Product Datasheets

Preparation and Storage

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

GENERAL ELISA PROTOCOL

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.

11 Citations: Showing 1 - 10
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  1. 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;20(1):6.
    Species: Human
    Sample Types: Urine
  2. 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;13(7):e0200537.
    Species: Human
    Sample Types: Plasma
  3. 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;80(0):5-13.
    Species: Human
    Sample Types: Platelets
  4. 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;0(0):.
    Species: Human
    Sample Types: Plasma
  5. Novel Antiplatelet Activity of Minocycline Involves Inhibition of MLK3-p38 Mitogen Activated Protein Kinase Axis
    Authors: Joseph W Jackson
    PLoS ONE, 2016;11(6):e0157115.
    Species: Human
    Sample Types: Plasma
  6. 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, 2014;370(5):433-43.
    Species: Human
    Sample Types: Plasma
  7. 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;117(14):3907-11.
    Species: Human
    Sample Types: Cell Culture Supernates
  8. 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;116(22):4703-11.
    Species: Human
    Sample Types: Cell Culture Supernates
  9. 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;151(5):495-503.
    Species: Human
    Sample Types: Plasma
  10. Normal ranges of angiogenesis regulatory proteins in human platelets.
    Authors: Peterson JE, Zurakowski D, Italiano JE
    Am. J. Hematol., 2010;85(7):487-93.
    Species: Human
    Sample Types: Cell Lysates
  11. 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;112(8):3154-63.
    Species: Human
    Sample Types: Cell Culture Supernates

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Reviews for Human CXCL4/PF4 DuoSet ELISA

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