Human CX3CL1/Fractalkine Quantikine ELISA Kit

  (15 citations)
(2 Reviews)
    
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Assay Procedure
Citations (15)
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  • 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 (100 uL), Serum (100 uL), EDTA Plasma (100 uL), Heparin Plasma (100 uL), Saliva (50 uL), Urine (50 uL), Human Milk (100 uL)
  • Sensitivity
    0.072 ng/mL
  • Assay Range
    0.2 - 10 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine, Human Milk)
  • Specificity
    Natural and recombinant human Fractalkine. This assay detects all forms of human Fractalkine containing the chemokine domain.
  • 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.
Control Available
QC93 , Quantikine Immunoassay Control Set 577 Human Fractalkine - Please Inquire
Product Summary
The Quantikine Human Fractalkine immunoassay is a 4.5 hour solid phase ELISA designed to measure Fractalkine in cell culture supernates, serum, plasma, saliva, urine, and human milk. It contains NS0-expressed recombinant human Fractalkine and has been shown to accurately quantitate the recombinant factor. Results obtained using natural Fractalkine 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 Fractalkine.

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, Saliva
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean 1.18 3.33 6.43 1.19 3.53 6.85
Standard Deviation 0.03 0.11 0.24 0.07 0.15 0.31
CV% 2.5 3.4 3.7 6.1 4.3 4.5

Serum, EDTA Plasma, Heparin Plasma, Urine, Human Milk
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean 1.24 3.8 7.12 1.23 3.59 6.88
Standard Deviation 0.04 0.08 0.17 0.11 0.2 0.37
CV% 3.2 2.1 2.4 8.9 5.6 5.4

Recovery

The recovery of Fractalkine spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 98 93-102
EDTA Plasma (n=4) 101 88-113
Heparin Plasma (n=4) 97 85-108
Serum (n=4) 99 88-111
Urine (n=4) 96 86-109
Linearity
To assess the linearity of the assay, samples containing or spiked with high concentrations of Fractalkine were diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
 CX3CL1/Fractalkine [HRP]
Product Datasheets

Certificate of Analysis Lookup

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Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: CX3CL1/Fractalkine
CX3CL1/Fractalkine is a transmembrane adhesion protein with a chemokine domain separated from the membrane by a mucin stalk. It is upregulated on many cell types during inflammation. The chemokine and mucin regions can be shed as a soluble chemokine that signals through the receptor CX3CR1. During extravasation, membrane-bound CX3CL1 traps leukocytes, then is cleaved to allow diapedesis. Soluble CX3CL1 protects neurons from microglial neurotoxicity, recruits macrophages for wound healing, recruits osteoclast precursors for bone resorption, and contributes to pathogenesis in coronary artery disease.
  • Entrez Gene IDs:
    6376 (Human); 20312 (Mouse); 89808 (Rat)
  • Alternate Names:
    ABCD-3; C3Xkine; chemokine (C-X3-C motif) ligand 1; CX3C membrane-anchored chemokine; CX3CL1; CXC3; CXC3C; FKN; Fractalkine; Neurotactin; neurotactin); NTNsmall inducible cytokine subfamily D (Cys-X3-Cys), member 1 (fractalkine; NTTSmall-inducible cytokine D1; SCYD1C-X3-C motif chemokine 1; small inducible cytokine subfamily D (Cys-X3-Cys), member-1
Related Research Areas
Assay Procedure
Refer to the product for complete assay procedure.

The conjugate must be kept cold during use. Bring all other 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. 100 µL Standard, Control, or Sample
  6.   Add 100 µL of Standard, control, or sample to each well within 15 minutes. Cover with a plate sealer, and incubate at 2-8 °C for 3 hours.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 200 µL cold Conjugate
  9.   Add 200 µL of cold Conjugate to each well. Cover with a new plate sealer, and incubate at 2-8 °C for 1 hour.
  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. 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:

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.

15 Citations: Showing 1 - 10
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Species
Sample Type
  1. Lipopolysaccharide-Binding Protein Downregulates Fractalkine through Activation of p38 MAPK and NF-?B
    Authors: X Huang, Y Zeng, Y Jiang, Y Qin, W Luo, S Xiang, SR Sooranna, L Pinhu
    Mediators Inflamm., 2017;2017(0):9734837.
    Species: Rat
    Sample Type: Serum
  2. Assessment of the Role of Metabolic Determinants on the Relationship between Insulin Sensitivity and Secretion
    PLoS ONE, 2016;11(12):e0168352.
    Species: Human
    Sample Type: Plasma
  3. Altered Monocyte and Endothelial Cell Adhesion Molecule Expression Is Linked to Vascular Inflammation in Human Immunodeficiency Virus Infection
    Authors: M Kulkarni, E Bowman, J Gabriel, T Amburgy, E Mayne, DA Zidar, C Maierhofer, AN Turner, JA Bazan, SL Koletar, MM Lederman, SF Sieg, NT Funderburg
    Open Forum Infect Dis, 2016;3(4):ofw224.
    Species: Human
    Sample Type: Plasma
  4. T lymphocytes and fractalkine contribute to myocardial ischemia/reperfusion injury in patients.
    Authors: Boag S, Das R, Shmeleva E, Bagnall A, Egred M, Howard N, Bennaceur K, Zaman A, Keavney B, Spyridopoulos I
    J Clin Invest, 2015;125(8):3063-76.
    Species: Human
    Sample Type: Serum
  5. Fractalkine-CX3CR1-dependent recruitment and retention of human CD1c+ myeloid dendritic cells by in vitro-activated proximal tubular epithelial cells.
    Authors: Kassianos A, Wang X, Sampangi S, Afrin S, Wilkinson R, Healy H
    Kidney Int, 2015;87(6):1153-63.
    Species: Human
    Sample Type: Tissue Homogenates
  6. Fractalkine is an independent predictor of mortality in patients with advanced heart failure.
    Thromb Haemost, 2012;108(6):1220-7.
    Species: Human
    Sample Type: Plasma
  7. Systemic biomarkers of inflammation and haemostasis in patients with chronic necrotizing pulmonary aspergillosis.
    BMC Infect Dis, 2012;12(0):144.
    Species: Human
    Sample Type: Serum
  8. Early detection of malignant pleural mesothelioma in asbestos-exposed individuals with a noninvasive proteomics-based surveillance tool.
    Authors: Ostroff R, Mehan M, Stewart A, Ayers D, Brody E, Williams S, Levin S, Black B, Harbut M, Carbone M, Goparaju C, Pass H
    PLoS ONE, 2012;7(10):e46091.
    Species: Human
    Sample Type: Serum
  9. Fractalkine is expressed in early and advanced atherosclerotic lesions and supports monocyte recruitment via CX3CR1.
    Authors: Stolla M, Pelisek J, von Bruhl M, Schafer A, Barocke V, Heider P, Lorenz M, Tirniceriu A, Steinhart A, Bauersachs J, Bray P, Massberg S, Schulz C
    PLoS ONE, 2012;7(8):e43572.
    Species: Human
    Sample Type: Serum
  10. Activated innate immunity and the involvement of CX3CR1-fractalkine in promoting hematuria in patients with IgA nephropathy.
    Kidney Int., 2012;82(5):548-60.
    Species: Human
    Sample Type: Urine
  11. Altered expression of CX3CL1 in patients with epilepsy and in a rat model.
    Authors: Xu Y, Zeng K, Han Y, Wang L, Chen D, Xi Z, Wang H, Wang X, Chen G
    Am. J. Pathol., 2012;180(5):1950-62.
    Species: Human
    Sample Type: CSF
  12. Identification of the chemokine CX3CL1 as a new regulator of malignant cell proliferation in epithelial ovarian cancer.
    Authors: Gaudin F, Nasreddine S, Donnadieu AC, Emilie D, Combadiere C, Prevot S, Machelon V, Balabanian K
    PLoS ONE, 2012;6(7):e21546.
    Species: Human
    Sample Type: Cell Culture Supernates
  13. Fractalkine is a novel human adipochemokine associated with type 2 diabetes.
    Authors: Shah R, Hinkle CC, Ferguson JF, Mehta NN, Li M, Qu L, Lu Y, Putt ME, Ahima RS, Reilly MP
    Diabetes, 2011;60(5):1512-8.
    Species: Human
    Sample Type: Plasma
  14. Characterization of synergistic induction of CX3CL1/fractalkine by TNF-alpha and IFN-gamma in vascular endothelial cells: an essential role for TNF-alpha in post-transcriptional regulation of CX3CL1.
    Authors: Matsumiya T, Ota K, Imaizumi T
    J. Immunol., 2010;184(8):4205-14.
    Species: Human
    Sample Type: Cell Culture Supernates
  15. Constitutive endocytosis of the chemokine CX3CL1 prevents its degradation by cell surface metalloproteases.
    Authors: Huang YW, Su P, Liu GY, Crow MR, Chaukos D, Yan H, Robinson LA
    J. Biol. Chem., 2009;284(43):29644-53.
    Species: Human
    Sample Type: Cell Culture Supernates
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Quantikine Immunoassay Control Set 577 Human Fractalkine

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Sample Tested H69AR human small cell lung cancer cell line
 Human CX3CL1/Fractalkine Quantikine ELISA Kit DCX310 Array DCX310
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 Human CX3CL1/Fractalkine Quantikine ELISA Kit DCX310
: Human CX3CL1/Fractalkine Quantikine ELISA Kit [DCX310].

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