Human CCL5/RANTES DuoSet ELISA

Catalog # Availability Size / Price Qty
DY278
DY278-05
Ancillary Products Available
Human CCL5 / RANTES ELISA Standard Curve
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Product Details
Procedure
Citations (40)
FAQs
Supplemental Products
Reviews (1)

Human CCL5/RANTES DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Sufficient Materials
For five or 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 CCL5/RANTES. 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 CCL5 / RANTES 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: CCL5/RANTES

CCL5/RANTES is a secreted beta-chemokine that plays a primary role in the inflammatory immune response through chemoattraction and activation of leukocytes that express CCR1, 3, 4, or 5. The two N-terminal residues of CCL5 can be removed by CD26/DPPIV, generating a protein that functions as a chemotaxis inhibitor. The deposition of CCL5 on activated vascular endothelial cells is crucial for monocyte adhesion to damaged vasculature. CCL5 is upregulated in breast cancer and promotes tumor progression through the attraction of proinflammatory macrophages in addition to its actions on tumor cells, stromal cells, and the vasculature. In humans, CCR5 binding to CCL5 inhibits the infectivity of R5 (M-tropic) but not X4 (T-tropic) strains of HIV-1.

Entrez Gene IDs:
6352 (Human); 20304 (Mouse); 403522 (Canine); 493689 (Feline)
Alternate Names:
CCL5; chemokine (C-C motif) ligand 5; D17S136Enormally T-expressed, and presumably secreted; EoCP; Eosinophil chemotactic cytokine; RANTES; SISd; SIS-delta; small inducible cytokine A5 (RANTES); small inducible cytokine subfamily A (Cys-Cys), member 5; Small-inducible cytokine A5; T cell-specific protein P228; T-cell specific protein p288; TCP228T-cell-specific protein RANTES

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 CCL5/RANTES 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.

40 Citations: Showing 1 - 10
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  1. IFI16 and cGAS cooperate in the activation of STING during DNA sensing in human keratinocytes
    Authors: JF Almine, CA O'Hare, G Dunphy, IR Haga, RJ Naik, A Atrih, DJ Connolly, J Taylor, IR Kelsall, AG Bowie, PM Beard, L Unterholzn
    Nat Commun, 2017;8(0):14392.
    Species: Human
    Sample Types: Cell Culture Supernates
  2. Pivotal role for the ESCRT-II complex subunit EAP30/SNF8 in IRF3-dependent innate antiviral defense
    Authors: K Kumthip, D Yang, NL Li, Y Zhang, M Fan, A Sethuraman, K Li
    PLoS Pathog., 2017;13(10):e1006713.
    Species: Human
    Sample Types: Cell Culture Supernates
  3. Zika Virus Persistently Infects and Is Basolaterally Released from Primary Human Brain Microvascular Endothelial Cells
    Authors: MC Mladinich, J Schwedes, ER Mackow
    MBio, 2017;8(4):.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. Malaria pigment stimulates chemokine production by human microvascular endothelium
    Authors: N Basilico, Y Corbett, S Alessandro, S Parapini, M Prato, D Girelli, P Misiano, P Olliaro, D Taramelli
    Acta Trop., 2017;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  5. Unfractionated Heparin Selectively Modulates the Expression of CXCL8, CCL2 and CCL5 in Endometrial Carcinoma Cells
    Authors: A Doster, U Schwarzig, M Zygmunt, J Rom, F Schütz, H Fluhr
    Anticancer Res, 2016;36(4):1535-44.
    Species: Human
    Sample Types: Cell Culture Supernates
  6. Forsythia suspensa Suppresses House Dust Mite Extract-Induced Atopic Dermatitis in NC/Nga Mice
    PLoS ONE, 2016;11(12):e0167687.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. Mitochondrial proteins NIP-SNAP-1 and -2 are a target for the immunomodulatory activity of clarithromycin, which involves NF-?B-mediated cytokine production
    Authors: Soh Yamamoto
    Biochem. Biophys. Res. Commun, 2016;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  8. Inflammatory and immunological profiles in patients with COPD: relationship with FEV 1 reversibility
    Authors: Cleriston Farias Queiroz
    J Bras Pneumol, 2016;42(4):241-247.
    Species: Human
    Sample Types: Serum
  9. Necroptosis suppresses inflammation via termination of TNF- or LPS-induced cytokine and chemokine production.
    Authors: Kearney C, Cullen S, Tynan G, Henry C, Clancy D, Lavelle E, Martin S
    Cell Death Differ, 2015;22(8):1313-27.
    Species: Human
    Sample Types: Cell Culture Supernates
  10. Role of hydrogen sulfide in paramyxovirus infections.
    Authors: Li H, Ma Y, Escaffre O, Ivanciuc T, Komaravelli N, Kelley J, Coletta C, Szabo C, Rockx B, Garofalo R, Casola A
    J Virol, 2015;89(10):5557-68.
    Species: Human
    Sample Types: Cell Culture Supernates
  11. Experimental hyperglycemia induces an increase of monocyte and T-lymphocyte content in adipose tissue of healthy obese women.
    Authors: Tencerova M, Kracmerova J, Krauzova E, Malisova L, Kovacova Z, Wedellova Z, Siklova M, Stich V, Rossmeislova L
    PLoS ONE, 2015;10(3):e0122872.
    Species: Human
    Sample Types: Plasma
  12. Resolving cancer-stroma interfacial signalling and interventions with micropatterned tumour-stromal assays.
    Authors: Shen K, Luk S, Hicks D, Elman J, Bohr S, Iwamoto Y, Murray R, Pena K, Wang F, Seker E, Weissleder R, Yarmush M, Toner M, Sgroi D, Parekkadan B
    Nat Commun, 2014;5(0):5662.
    Species: Human
    Sample Types: Cell Culture Supernates
  13. Proteome-wide analysis and CXCL4 as a biomarker in systemic sclerosis
    Authors: L van Bon, AJ Affandi, J Broen, RB Christmann, RJ Marijnisse, L Stawski, GA Farina, G Stifano, AL Mathes, M Cossu, M York, C Collins, M Wenink, R Huijbens, R Hesselstra, T Saxne, M DiMarzio, D Wuttge, SK Agarwal, JD Reveille, S Assassi, M Mayes, Y Deng, JP Drenth, J de Graaf, M den Heijer, CG Kallenberg, M Bijl, A Loof, WB van den Be, LA Joosten, V Smith, F de Keyser, R Scorza, C Lunardi, PL van Riel, M Vonk, W van Heerde, S Meller, B Homey, L Beretta, M Roest, M Trojanowsk, R Lafyatis, TR Radstake
    N. Engl. J. Med, 2014;370(5):433-43.
    Species: Human
    Sample Types: Plasma
  14. Controlled intramyocardial release of engineered chemokines by biodegradable hydrogels as a treatment approach of myocardial infarction.
    Authors: Projahn D, Simsekyilmaz S, Singh S, Kanzler I, Kramp B, Langer M, Burlacu A, Bernhagen J, Klee D, Zernecke A, Hackeng T, Groll J, Weber C, Liehn E, Koenen R
    J Cell Mol Med, 2014;18(5):790-800.
    Species: Human
    Sample Types: Cell Culture Supernates
  15. Human osteoarthritic cartilage shows reduced in vivo expression of IL-4, a chondroprotective cytokine that differentially modulates IL-1beta-stimulated production of chemokines and matrix-degrading enzymes in vitro.
    Authors: Assirelli E, Pulsatelli L, Dolzani P, Platano D, Olivotto E, Filardo G, Trisolino G, Facchini A, Borzi R, Meliconi R
    PLoS ONE, 2014;9(5):e96925.
    Species: Human
    Sample Types: Cell Culture Supernates
  16. The small GTPase Arf6 is essential for the Tram/Trif pathway in TLR4 signaling.
    Authors: Van Acker T, Eyckerman S, Vande Walle L, Gerlo S, Goethals M, Lamkanfi M, Bovijn C, Tavernier J, Peelman F
    J Biol Chem, 2014;289(3):1364-76.
    Species: Human
    Sample Types: Cell Culture Supernates
  17. Characterisation of P2Y(12) receptor responsiveness to cysteinyl leukotrienes.
    Authors: Foster H, Fuerst E, Lee T, Cousins D, Woszczek G
    PLoS ONE, 2013;8(3):e58305.
    Species: Human
    Sample Types: Plasma
  18. Intradermal delivery of TLR agonists in a human explant skin model: preferential activation of migratory dendritic cells by polyribosinic-polyribocytidylic acid and peptidoglycans.
    Authors: Oosterhoff D, Heusinkveld M, Lougheed S, Kosten I, Lindstedt M, Bruijns S, van Es T, van Kooyk Y, van der Burg S, de Gruijl T
    J Immunol, 2013;190(7):3338-45.
    Species: Human
    Sample Types: Cell Culture Supernates
  19. Human cytomegalovirus induces a biphasic inflammatory response in primary endothelial cells.
    Authors: Jeffery H, Soderberg-Naucler C, Butler L
    J Virol, 2013;87(11):6530-5.
    Species: Human
    Sample Types: Cell Culture Supernates
  20. 1,25-dihydroxyvitamin D3 protects against macrophage-induced activation of NFkappaB and MAPK signalling and chemokine release in human adipocytes.
    Authors: Ding C, Wilding J, Bing C
    PLoS ONE, 2013;8(4):e61707.
    Species: Human
    Sample Types: Cell Culture Supernates
  21. Measles virus causes immunogenic cell death in human melanoma.
    Authors: Donnelly, O G, Errington-Mais, F, Steele, L, Hadac, E, Jennings, V, Scott, K, Peach, H, Phillips, R M, Bond, J, Pandha, H, Harrington, K, Vile, R, Russell, S, Selby, P, Melcher, A A
    Gene Ther, 2013;20(1):7-15.
    Species: Human
    Sample Types: Cell Culture Supernates
  22. IRF5 promotes inflammatory macrophage polarization and T(H)1-T(H)17 responses.
    Authors: Krausgruber T, Blazek K, Smallie T
    Nat. Immunol., 2011;12(3):231-8.
    Species: Human
    Sample Types: Cell Culture Supernates
  23. IL-17 amplifies human contact hypersensitivity by licensing hapten nonspecific Th1 cells to kill autologous keratinocytes.
    Authors: Pennino D, Eyerich K, Scarponi C, Carbone T, Eyerich S, Nasorri F, Garcovich S, Traidl-Hoffmann C, Albanesi C, Cavani A
    J. Immunol., 2010;184(9):4880-8.
    Species: Human
    Sample Types: Cell Culture Supernates
  24. A role for the human nucleotide-binding domain, leucine-rich repeat-containing family member NLRC5 in antiviral responses.
    Authors: Neerincx A, Lautz K, Menning M, Kremmer E, Zigrino P, Hosel M, Buning H, Schwarzenbacher R, Kufer TA
    J. Biol. Chem., 2010;285(34):26223-32.
    Species: Human
    Sample Types: Cell Culture Supernates
  25. Thrombocytopenia in early malaria is associated with GP1b shedding in absence of systemic platelet activation and consumptive coagulopathy.
    Authors: de Mast Q, de Groot PG, van Heerde WL, Roestenberg M, van Velzen JF, Verbruggen B, Roest M, McCall M, Nieman AE, Westendorp J, Syafruddin D, Fijnheer R, van Dongen-Lases EC, Sauerwein RW, van der Ven AJ
    Br. J. Haematol., 2010;151(5):495-503.
    Species: Human
    Sample Types: Plasma
  26. Novel markers of inflammation identified in tumor necrosis factor receptor-associated periodic syndrome (TRAPS) by transcriptomic analysis of effects of TRAPS-associated tumor necrosis factor receptor type I mutations in an endothelial cell line.
    Authors: Rebelo SL, Amel-Kashipaz MR, Radford PM, Bainbridge SE, Fiets R, Fang J, McDermott EM, Powell RJ, Todd I, Tighe PJ
    Arthritis Rheum., 2009;60(1):269-80.
    Species: Human
    Sample Types: Cell Culture Supernates
  27. 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
  28. Expanded-polyglutamine huntingtin protein suppresses the secretion and production of a chemokine (CCL5/RANTES) by astrocytes.
    Authors: Chou SY, Weng JY, Lai HL, Liao F, Sun SH, Tu PH, Dickson DW, Chern Y
    J. Neurosci., 2008;28(13):3277-90.
    Species: Human
    Sample Types: Cell Culture Supernates
  29. Altered levels of CC chemokines during pulmonary CMV predict BOS and mortality post-lung transplantation.
    Authors: Weigt SS, Elashoff RM, Keane MP, Strieter RM, Gomperts BN, Xue YY, Ardehali A, Gregson AL, Kubak B, Fishbein MC, Saggar R, Ross DJ, Lynch JP, Zisman DA, Belperio JA
    Am. J. Transplant., 2008;8(7):1512-22.
    Species: Human
    Sample Types: BALF
  30. CD8 chemokine receptors in chronic obstructive pulmonary disease.
    Authors: Smyth LJ, Starkey C, Gordon FS, Vestbo J, Singh D
    Clin. Exp. Immunol., 2008;154(1):56-63.
    Species: Human
    Sample Types: BALF
  31. Preadipocyte response and impairment of differentiation in an inflammatory environment.
    Authors: Poulain-Godefroy O, Froguel P
    Biochem. Biophys. Res. Commun., 2007;356(3):662-7.
    Species: Human
    Sample Types: Cell Culture Supernates
  32. PAPP-A and osteoprotegerin, together with interleukin-8 and RANTES, are elevated in the peritoneal fluid of women with endometriosis.
    Authors: Bersinger NA, von Roten S, Wunder DM, Raio L, Dreher E, Mueller MD
    Am. J. Obstet. Gynecol., 2006;195(1):103-8.
    Species: Human
    Sample Types: Peritoneal Fluid
  33. The cytokine IL-1beta activates IFN response factor 3 in human fetal astrocytes in culture.
    Authors: Rivieccio MA, John GR, Song X, Suh HS, Zhao Y, Lee SC, Brosnan CF
    J. Immunol., 2005;174(6):3719-26.
    Species: Human
    Sample Types: Cell Culture Supernates
  34. Manipulation of host cytokine network by ticks: a potential gateway for pathogen transmission.
    Authors: Hajnicka V, Vancova I, Kocakova P, Slovak M, Gasperik J, Slavikova M, Hails RS, Labuda M, Nuttall PA
    Parasitology, 2005;130(0):333-42.
    Species: Human
    Sample Types: Saliva
  35. Rapid chemokine secretion from endothelial cells originates from 2 distinct compartments.
    Authors: Oynebraten I, Bakke O, Brandtzaeg P, Johansen FE, Haraldsen G
    Blood, 2004;104(2):314-20.
    Species: Human
    Sample Types: Cell Culture Supernates
  36. Involvement of Fc gamma RI (CD64) in the mechanism of HIV-1 inhibition by polyclonal IgG purified from infected patients in cultured monocyte-derived macrophages.
    Authors: Holl V, Hemmerter S, Burrer R, Schmidt S, Bohbot A, Aubertin AM, Moog C
    J. Immunol., 2004;173(10):6274-83.
    Species: Human
    Sample Types: Cell Culture Supernates
  37. Chemokine secretion of rheumatoid arthritis synovial fibroblasts stimulated by Toll-like receptor 2 ligands.
    Authors: Pierer M, Rethage J, Seibl R, Lauener R, Brentano F, Wagner U, Hantzschel H, Michel BA, Gay RE, Gay S, Kyburz D
    J. Immunol., 2004;172(2):1256-65.
    Species: Human
    Sample Types: Cell Culture Supernates
  38. Virally stimulated plasmacytoid dendritic cells produce chemokines and induce migration of T and NK cells.
    Authors: Megjugorac NJ, Young HA, Amrute SB, Olshalsky SL, Fitzgerald-Bocarsly P
    J. Leukoc. Biol., 2004;75(3):504-14.
    Species: Human
    Sample Types: Cell Culture Supernates
  39. Phenotypic and functional analysis of T cells homing into the CSF of subjects with inflammatory diseases of the CNS.
    Authors: Giunti D, Borsellino G, Benelli R, Marchese M, Capello E, Valle MT, Pedemonte E, Noonan D, Albini A, Bernardi G, Mancardi GL, Battistini L, Uccelli A
    J. Leukoc. Biol., 2003;73(5):584-90.
    Species: Human
    Sample Types: CSF
  40. Differential production of RANTES and MCP-1 in synovial fluid from the inflamed human knee.
    Authors: Conti P, Reale M, Barbacane RC, Castellani ML, Orso C
    Immunol. Lett., 2002;80(2):105-11.
    Species: Human
    Sample Types: Serum

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Human CCL5/RANTES DuoSet ELISA
By Anonymous on 07/17/2018
Sample Tested: EDTA Plasma