Human CX3CL1/Fractalkine DuoSet ELISA

Catalog # Availability Size / Price Qty
DY365
Ancillary Products Available
CX3CL1/Fractalkine ELISA Kit ELISA
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Citations (19)
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Human CX3CL1/Fractalkine 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 CX3CL1/Fractalkine. 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 CX3CL1 / Fractalkine 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: 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

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 CX3CL1/Fractalkine 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.

19 Citations: Showing 1 - 10
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  1. ADAM-17 is expressed on rheumatoid arthritis fibroblast-like synoviocytes and regulates proinflammatory mediator expression and monocyte adhesion
    Authors: S Ishii, T Isozaki, H Furuya, H Takeuchi, Y Tsubokura, K Inagaki, T Kasama
    Arthritis Res. Ther., 2018;20(1):159.
    Species: Human
    Sample Types: Serum
  2. Glycogen synthase kinase-3? regulates fractalkine production by altering its trafficking from Golgi to plasma membrane: implications for Alzheimer's disease
    Authors: Jesús Ávila
    Cell. Mol. Life Sci., 2016;0(0):.
    Species: Human
    Sample Types: Tissue Homogenates
  3. Serum level of soluble CX3CL1/fractalkine is elevated in patients with polymyositis and dermatomyositis, which is correlated with disease activity.
    Authors: Suzuki F, Kubota T
    Arthritis Res. Ther., 2012;14(2):R48.
    Species: Human
    Sample Types: Serum
  4. CX3CL1 expression in the conjunctiva is involved in immune cell trafficking during toxic ocular surface inflammation.
    Authors: Denoyer A, Godefroy D, Celerier I, Frugier J, Riancho L, Baudouin F, Rostene W, Baudouin C
    Mucosal Immunol, 2012;5(0):702.
    Species: Human
    Sample Types: Cell Culture Supernates
  5. Thromboxane prostanoid receptor stimulation induces shedding of the transmembrane chemokine CX3CL1 yet enhances CX3CL1-dependent leukocyte adhesion.
    Authors: Tole S, Durkan AM, Huang YW, Liu GY, Leung A, Jones LL, Taylor JA, Robinson LA
    Am. J. Physiol., Cell Physiol., 2010;298(6):C1469-80.
    Species: Human
    Sample Types: Cell Lysates
  6. Expression of ADAM-17, TIMP-3 and fractalkine in the human adult brain endothelial cell line, hCMEC/D3, following pro-inflammatory cytokine treatment.
    Authors: Hurst LA, Bunning RA, Couraud PO, Romero IA, Weksler BB, Sharrack B, Woodroofe MN
    J. Neuroimmunol., 2009;210(1):108-12.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. The involvement of the fractalkine receptor in the transmigration of neuroblastoma cells through bone-marrow endothelial cells.
    Authors: Nevo I, Sagi-Assif O, Meshel T, Ben-Baruch A, Johrer K, Greil R, Trejo LE, Kharenko O, Feinmesser M, Yron I, Witz IP
    Cancer Lett., 2009;273(1):127-39.
    Species: Human
    Sample Types: Cell Culture Supernates
  8. CX3CR1 is expressed by prostate epithelial cells and androgens regulate the levels of CX3CL1/fractalkine in the bone marrow: potential role in prostate cancer bone tropism.
    Authors: Jamieson WL, Shimizu S, D'Ambrosio JA, Meucci O, Fatatis A
    Cancer Res., 2008;68(6):1715-22.
    Species: Human
    Sample Types: Tissue Secretion
  9. Urinary fractalkine is a marker of acute rejection.
    Authors: Peng W, Chen J, Jiang Y, Wu J, Shou Z, He Q, Wang Y, Chen Y, Wang H
    Kidney Int., 2008;74(11):1454-60.
    Species: Human
    Sample Types: Urine
  10. Vitamin D and glucocorticoids differentially modulate chemokine expression in human airway smooth muscle cells.
    Authors: Banerjee A, Damera G, Bhandare R, Gu S, Lopez-Boado Y, Panettieri R, Tliba O
    Br. J. Pharmacol., 2008;155(1):84-92.
    Species: Human
    Sample Types: Cell Culture Supernates
  11. Cytokine expression profiling of the myocardium reveals a role for CX3CL1 (fractalkine) in heart failure.
    Authors: Husberg C, Nygard S, Finsen AV, Damas JK, Frigessi A, Oie E, Waehre A, Gullestad L, Aukrust P, Yndestad A, Christensen G
    J. Mol. Cell. Cardiol., 2008;45(2):261-9.
    Species: Human
    Sample Types: Serum
  12. Proteomics analysis of Hodgkin lymphoma: identification of new players involved in the cross-talk between HRS cells and infiltrating lymphocytes.
    Authors: Ma Y, Visser L, Roelofsen H, de Vries M, Diepstra A, van Imhoff G, van der Wal T, Luinge M, Alvarez-Llamas G, Vos H, Poppema S, Vonk R, Van Den Berg A
    Blood, 2007;111(4):2339-46.
    Species: Human
    Sample Types: Cell Culture Supernates
  13. Endothelial damage from cytomegalovirus-specific host immune response can be prevented by targeted disruption of fractalkine-CX3CR1 interaction.
    Authors: Bolovan-Fritts CA, Spector SA
    Blood, 2007;111(1):175-82.
    Species: Human
    Sample Types: Cell Culture Supernates
  14. High T-cell response to human cytomegalovirus induces chemokine-mediated endothelial cell damage.
    Authors: Bolovan-Fritts CA, Trout RN, Spector SA
    Blood, 2007;110(6):1857-63.
    Species: Human
    Sample Types: Cell Culture Supernates
  15. The uremic solute p-cresol decreases leukocyte transendothelial migration in vitro.
    Authors: Faure V, Cerini C, Paul P, Berland Y, Dignat-George F, Brunet P
    Int. Immunol., 2006;18(10):1453-9.
    Species: Human
    Sample Types: Cell Culture Supernates
  16. Soluble fractalkine in the cerebrospinal fluid of patients with neuropsychiatric lupus.
    Authors: Sato E, Iikuni N, Yoshio T, Minota S, Kamatani N, Okamoto H
    Ann. Rheum. Dis., 2006;65(9):1257-9.
    Species: Human
    Sample Types: CSF
  17. Fractalkine and CX3CR1 are involved in the recruitment of intraepithelial lymphocytes of intrahepatic bile ducts.
    Authors: Isse K, Harada K, Zen Y, Kamihira T, Shimoda S, Harada M, Nakanuma Y
    Hepatology, 2005;41(3):506-16.
    Species: Human
    Sample Types: Serum
  18. Fractalkine reduces N-methyl-d-aspartate-induced calcium flux and apoptosis in human neurons through extracellular signal-regulated kinase activation.
    Authors: Deiva K, Geeraerts T, Salim H, Leclerc P, Hery C, Hugel B, Freyssinet JM, Tardieu M
    Eur. J. Neurosci., 2004;20(12):3222-32.
    Species: Human
    Sample Types: Cell Culture Supernates
  19. Human cytomegalovirus-specific CD4+-T-cell cytokine response induces fractalkine in endothelial cells.
    Authors: Bolovan-Fritts CA, Trout RN, Spector SA
    J. Virol., 2004;78(23):13173-81.
    Species: Human
    Sample Types: Cell Culture Supernates

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