Overview

  • Product name

    Mouse PF4 ELISA Kit (CXCL4)
    See all PF4 kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Mouse serum 8 3.2%
    Inter-assay
    Sample n Mean SD CV%
    Mouse serum 3 5.5%
  • Sample type

    Cell culture supernatant, Serum, EDTA Plasma, Citrate Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    148 pg/ml
  • Range

    0.25 ng/ml - 16 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 97 93% - 99%
    Cell culture media 91 83% - 96%
    EDTA Plasma 101 100% - 102%
    Citrate Plasma 110 106% - 114%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Mouse
    Does not react with: Rat, Sheep, Rabbit, Guinea pig, Cow, Dog, Human, Pig, Syrian hamster
  • Product overview

    Abcam’s mouse PF4 (CXCL4) in vitro SimpleStep ELISA™ (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of mouse PF4 protein in serum, plasma and cell culture supernatant samples.

    The SimpleStep ELISA™ employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.

  • Notes

    Platelet factor 4 (PF4) is a small cytokine belonging to the CXC chemokine family that is also known as chemokine (C-X-C motif) ligand 4 (CXCL4). Chemokines play fundamental roles in the development, homeostasis, and function of the immune system, and they have effects on cells of the central nervous system as well as on endothelial cells involved in angiogenesis and angiostasis. PF4 is a 70-amino acid protein (8 kDa) that is released from the alpha-granules of activated platelets and binds with high affinity to heparin. Specifically, the active protein is a tetramer of PF4 subunits that forms a ring of heparin-binding positive charges from sites at the C-terminal region of each monomer. In contrast to other CXC chemokines, PF4 does not contain an ELR motif and lacks binding to nearly all chemokine receptors. Mouse PF4 shares 64% and 89% amino acid identity with human and rat PF4, respectively.

    As a strong chemoattractant for neutrophils and fibroblasts, PF4 probably has a role in inflammation and wound repair. However, the major physiologic role of PF4 appears to be neutralization of heparin-like molecules on the endothelial surface of blood vessels, thereby inhibiting local antithrombin III activity and promoting coagulation. There can be over 100-fold higher concentrations of PF4 within clots than within normal serum. Therapeutic doses of heparin (an anticoagulant) neutralize PF4 pro-coagulant effects. The complex between heparin and PF4 can be immunogenic, producing the pathological syndrome HITT (heparin-induced thrombocytopenia and thrombosis). In addition, immunogenic complexes of PF4 with apolipoprotein H can contribute to antiphospholipid syndrome (APS).

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate (12 x 8 well strips)

Properties

Associated products

Applications

Our Abpromise guarantee covers the use of ab202403 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • Background-subtracted data values (mean +/- SD) are graphed.

  • Native mouse PF4 protein was measured in serum (diluted 1:2,500 in Sample Diluent NS) in a 2-fold dilution series in Sample Diluent NS. The interpolated dilution factor corrected values are graphed (mean +/- SD).

  • Native mouse PF4 protein was measured in citrate plasma (1:300), EDTA plasma (1:400), and Platelet Poor EDTA plasma (1:350) in a 2-fold dilution series in Sample Diluent NS. The interpolated dilution factor corrected values are graphed (mean +/- SD).

  • J774A.1 cells were cultured in HGDMEM with 10% fetal calf serum, and 100 µg/mL of Kanamycin. J774A.1 cells were treated for 72 hours in the presence and absence of 1.5% PHA and 10ng/mL of PMA. Samples were loaded on the plate diluted 1:10 and the concentrations of mouse PF4 were interpolated from a calibration curve diluted in Sample Diluent NS. The interpolated dilution factor corrected values are graphed (mean +/- SD).

Protocols

References

ab202403 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

There are currently no Customer reviews or Questions for ab202403.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up