Mouse CXCL4/PF4 Quantikine ELISA Kit

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MCX400
Mouse CXCL4/PF4 ELISA Standard Curve
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Citations (8)
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Mouse 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
0.01 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
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 0.23 0.41 1.51 0.25 0.45 1.46
Standard Deviation 0.013 0.029 0.046 0.019 0.035 0.11
CV% 5.7 7.1 3 7.6 7.8 7.5

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 ELISA Linearity

Scientific Data

Mouse 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
&#9888; 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. 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 for Mouse 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.

8 Citations: Showing 1 - 8
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  1. Perivascular niche cells sense thrombocytopenia and activate hematopoietic stem cells in an IL-1 dependent manner
    Authors: Luis, TC;Barkas, N;Carrelha, J;Giustacchini, A;Mazzi, S;Norfo, R;Wu, B;Aliouat, A;Guerrero, JA;Rodriguez-Meira, A;Bouriez-Jones, T;Macaulay, IC;Jasztal, M;Zhu, G;Ni, H;Robson, MJ;Blakely, RD;Mead, AJ;Nerlov, C;Ghevaert, C;Jacobsen, SEW;
    Nature communications
    Species: Mouse
    Sample Types: Bone Marrow Fluid
  2. Platelet factors attenuate inflammation and rescue cognition in ageing
    Authors: Schroer, AB;Ventura, PB;Sucharov, J;Misra, R;Chui, MKK;Bieri, G;Horowitz, AM;Smith, LK;Encabo, K;Tenggara, I;Couthouis, J;Gross, JD;Chan, JM;Luke, A;Villeda, SA;
    Nature
    Species: Mouse
    Sample Types: Plasma
  3. Platelet-derived chemokines promote skeletal muscle regeneration by guiding neutrophil recruitment to injured muscles
    Authors: Graca, FA;Stephan, A;Minden-Birkenmaier, BA;Shirinifard, A;Wang, YD;Demontis, F;Labelle, M;
    Nature communications
    Species: Mouse
    Sample Types: Plasma
  4. High ploidy large cytoplasmic megakaryocytes are hematopoietic stem cells regulators and essential for platelet production
    Authors: SY Heazlewood, T Ahmad, B Cao, H Cao, M Domingues, X Sun, CK Heazlewood, S Li, B Williams, M Fulton, JF White, T Nebl, CM Nefzger, JM Polo, BT Kile, F Kraus, MT Ryan, YB Sun, PFM Choong, SL Ellis, ML Anko, SK Nilsson
    Nature Communications, 2023-04-13;14(1):2099.
    Species: Mouse
    Sample Types: Plasma
  5. CD8+ T cells inhibit metastasis and CXCL4 regulates its function
    Authors: R Joseph, R Soundarara, S Vasaikar, F Yang, KL Allton, L Tian, P den Hollan, S Isgandarov, M Haemmerle, B Mino, T Zhou, C Shin, M Martinez-P, AA Sahin, J Rodriguez-, J Gelovani, JT Chang, G Acharya, AK Sood, II Wistuba, DL Gibbons, LM Solis, MC Barton, N Varadaraja, JM Rosen, XH Zhang, SA Mani
    British Journal of Cancer, 2021-04-01;0(0):.
    Species: Mouse
    Sample Types: Cell Lysates
  6. 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, 2017-08-21;7(1):8402.
    Species: Mouse
    Sample Types: Plasma
  7. 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, 2015-08-20;10(8):e0135738.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  8. 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, 2015-03-20;10(3):e0119739.
    Species: Mouse
    Sample Types: Cell Culture Supernates

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