Mouse CX3CL1/Fractalkine Quantikine ELISA Kit

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MCX310
Mouse CX3CL1/Fractalkine ELISA Standard Curve
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Citations (8)
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Mouse CX3CL1/Fractalkine 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), Tissue Homogenates (50 uL), Serum (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL)
Sensitivity
0.32 ng/mL
Assay Range
0.6 - 40 ng/mL (Cell Culture Supernates, Tissue Homogenates, Serum, EDTA Plasma, Heparin Plasma)
Specificity
Natural and recombinant mouse Fractalkine (chemokine and extracellular domains)
Cross-reactivity
Cross-reactivity observed with 1 or more 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 Fractalkine Immunoassay is a 4.5 hour solid-phase ELISA designed to measure mouse Fractalkine in cell culture supernates, tissue homogenates, mouse serum, and plasma. It contains E. coli-expressed recombinant mouse Fractalkine chemokine domain and antibodies raised against the recombinant protein. Results obtained for naturally occurring soluble mouse Fractalkine containing the chemokine domain and the mucin stalk domain 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 natural mouse 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, Tissue Homogenates, Serum, EDTA Plasma, Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 41 41 41
Mean 1.9 5.2 18.6 1.4 5.5 20
Standard Deviation 0.19 0.42 1.7 0.17 0.31 0.92
CV% 10 8.1 9.1 12.1 5.6 4.6

Recovery

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

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=5) 101 94-106
EDTA Plasma (n=6) 95 83-109
Heparin Plasma (n=6) 95 81-110
Serum (n=6) 98 91-107
Tissue Homogenates (n=1) 100 100

Linearity

To assess the linearity of the assay, samples containing or spiked with high concentrations of mouse Fractalkine in each matrix were diluted with Calibrator Diluent and assayed.
Mouse CX3CL1/Fractalkine ELISA Linearity

Data Examples

Mouse CX3CL1/Fractalkine ELISA Standard Curve

Product Datasheets

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: 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
⚠ 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 CX3CL1/Fractalkine 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. Sex differences in microglial CX3CR1 signalling determine obesity susceptibility in mice
    Authors: MD Dorfman, JE Krull, JD Douglass, R Fasnacht, F Lara-Lince, TH Meek, X Shi, V Damian, HT Nguyen, ME Matsen, GJ Morton, JP Thaler
    Nat Commun, 2017;8(0):14556.
    Species: Mouse
    Sample Types: Tissue Homogenates
  2. Intravenous hMSCs Ameliorate Acute Pancreatitis in Mice via Secretion of Tumor Necrosis Factor-? Stimulated Gene/Protein 6
    Sci Rep, 2016;6(0):38438.
    Species: Mouse
    Sample Types: Serum
  3. Fibroblast growth factor receptor 1 activation in mammary tumor cells promotes macrophage recruitment in a CX3CL1-dependent manner.
    Authors: Reed, Johanna, Stone, Matthew, Beadnell, Thomas C, Ryu, Yungil, Griffin, Timothy, Schwertfeger, Kathryn
    PLoS ONE, 2012;7(9):e45877.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  4. Ozone inhalation promotes CX3CR1-dependent maturation of resident lung macrophages that limit oxidative stress and inflammation.
    Authors: Tighe RM, Li Z, Potts EN, Frush S, Liu N, Gunn MD, Foster WM, Noble PW, Hollingsworth JW
    J. Immunol., 2011;187(9):4800-8.
    Species: Mouse
    Sample Types: BALF
  5. A novel mechanism for CCR4 in the regulation of macrophage activation in bleomycin-induced pulmonary fibrosis.
    Authors: Trujillo G, O'Connor EC, Kunkel SL, Hogaboam CM
    Am. J. Pathol., 2008;172(5):1209-21.
    Species: Mouse
    Sample Types: Tissue Homogenates
  6. Essential involvement of CX3CR1-mediated signals in the bactericidal host defense during septic peritonitis.
    Authors: Ishida Y, Hayashi T, Goto T, Kimura A, Akimoto S, Mukaida N, Kondo T
    J. Immunol., 2008;181(6):4208-18.
    Species: Mouse
    Sample Types: Peritoneal Lavage Fluid
  7. Down-regulation of surface fractalkine by RNA interference in B16 melanoma reduced tumor growth in mice.
    Authors: Ren T, Chen Q, Tian Z, Wei H
    Biochem. Biophys. Res. Commun., 2007;364(4):978-84.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  8. The arthritis severity quantitative trait loci Cia4 and Cia6 regulate neutrophil migration into inflammatory sites and levels of TNF-alpha and nitric oxide.
    Authors: Laragione T, Yarlett NC, Brenner M, Mello A, Sherry B, Miller EJ, Metz CN, Gulko PS
    J. Immunol., 2007;178(4):2344-51.
    Species: Rat
    Sample Types: Air Pouch Exudate

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