Human CCL19/MIP-3 beta DuoSet ELISA

Catalog # Availability Size / Price Qty
DY361
Ancillary Products Available
Human CCL19 / MIP-3 beta ELISA Standard Curve
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Product Details
Procedure
Citations (19)
FAQs
Supplemental Products
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Human CCL19/MIP-3 beta 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 CCL19/MIP-3beta. 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.

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

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 CCL19 / MIP-3 beta 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.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: CCL19/MIP-3 beta

Macrophage inflammatory protein 3 beta (MIP-3 beta), also known as ELC (EBI1-ligand chemokine) and exodus-3, is constitutively expressed in various lymphoid tissues, including thymus, lymph nodes, appendix and spleen. It is distantly related to other CC chemokines (20-30% amino acid sequence identity).

Entrez Gene IDs:
6363 (Human); 24047 (Mouse); 362506 (Rat)
Alternate Names:
beta chemokine exodus-3; Beta-chemokine exodus-3; CC chemokine ligand 19; C-C motif chemokine 19; CCL19; chemokine (C-C motif) ligand 19; CKb11; EBI1-ligand chemokine; ELC; ELCMIP-3-beta; Epstein-Barr virus-induced molecule 1 ligand chemokine; exodus-3; Macrophage inflammatory protein 3 beta; macrophage inflammatory protein 3-beta; MGC34433; MIP3 beta; MIP-3 beta; MIP-3b; MIP3BCK beta-11; SCYA19EBI1 ligand chemokine; small inducible cytokine subfamily A (Cys-Cys), member 19; Small-inducible cytokine A19

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 CCL19/MIP-3 beta 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. Altered B lymphocyte homeostasis and functions in systemic sclerosis
    Authors: A Forestier, T Guerrier, M Jouvray, J Giovannell, G Lefèvre, V Sobanski, C Hauspie, E Hachulla, PY Hatron, H Zephir, P Vermersch, M Labalette, D Launay, S Dubucquoi
    Autoimmun Rev, 2018;0(0):.
    Species: Human
    Sample Types: Serum
    Applications: ELISA Capture
  2. Distinctive expression of T cell guiding molecules in human autoimmune lymph node stromal cells upon TLR3 triggering
    Authors: JS Hähnlein, TH Ramwadhdoe, JF Semmelink, IY Choi, FH Berger, M Maas, DM Gerlag, PP Tak, TBH Geijtenbee, LGM van Baarse
    Sci Rep, 2018;8(1):1736.
    Species: Human
    Sample Types: Cell Culture Supernates
  3. Stimulation of osteoclast migration and bone resorption by C-C chemokine ligands 19 and 21
    Authors: J Lee, C Park, HJ Kim, YD Lee, ZH Lee, YW Song, HH Kim
    Exp. Mol. Med., 2017;49(7):e358.
    Species: Human
    Sample Types: Serum
  4. Human Lymph Node-Derived Fibroblastic and Double-Negative Reticular Cells Alter Their Chemokines and Cytokines Expression Profile Following Inflammatory Stimuli
    Authors: P Severino, DT Palomino, H Alvarenga, CB Almeida, DC Pasqualim, A Cury, PR Salvalaggi, AL De Vasconc, MC Andrade, T Aloia, S Bromberg, LV Rizzo, FA Rocha, LC Marti
    Front Immunol, 2017;8(0):141.
    Species: Human
    Sample Types: Cell Culture Supernates
  5. Cytokine network in scrub typhus: high levels of interleukin-8 are associated with disease severity and mortality.
    Authors: Astrup E, Janardhanan J, Otterdal K, Ueland T, Prakash J, Lekva T, Strand O, Abraham O, Thomas K, Damas J, Mathews P, Mathai D, Aukrust P, Varghese G
    PLoS Negl Trop Dis, 2014;8(2):e2648.
    Species: Human
    Sample Types: Plasma
  6. Induction of immunoregulatory CD271+ cells by metastatic tumor cells that express human endogenous retrovirus H.
    Authors: Kudo-Saito C, Yura M, Yamamoto R, Kawakami Y
    Cancer Res, 2014;74(5):1361-70.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. Feasibility of the use of combinatorial chemokine arrays to study blood and CSF in multiple sclerosis.
    Authors: Edwards, Keith R, Goyal, Jaya, Plavina, Tatiana, Czerkowicz, Julie, Goelz, Susan, Ranger, Ann, Cadavid, Diego, Browning, Jeffrey
    PLoS ONE, 2013;8(11):e81007.
    Species: Human
    Sample Types: Serum
  8. Alveolar fibrocyte percentage is an independent predictor of poor outcome in patients with acute lung injury.
    Authors: Quesnel C, Piednoir P, Gelly J, Nardelli L, Garnier M, Lecon V, Lasocki S, Bouadma L, Philip I, Elbim C, Mentre F, Soler P, Crestani B, Dehoux M
    Crit. Care Med., 2012;40(1):21-8.
    Species: Human
    Sample Types: BALF
  9. Adenovirus-engineered human dendritic cells induce natural killer cell chemotaxis via CXCL8/IL-8 and CXCL10/IP-10.
    Authors: Vujanovic, Lazar, Ballard, Wenners, Thorne, Stephen, Vujanovic, Nikola L, Butterfield, Lisa H
    Oncoimmunology, 2012;1(4):448-457.
    Species: Human
    Sample Types: Cell Culture Supernates
  10. Identification of a New Phenotype of Tolerogenic Human Dendritic Cells Induced by Fungal Proteases from Aspergillus oryzae.
    Authors: Zimmer A, Luce S, Gaignier F, Nony E, Naveau M, Biola-Vidamment A, Pallardy M, Van Overtvelt L, Mascarell L, Moingeon P
    J. Immunol., 2011;186(7):3966-76.
    Species: Human
    Sample Types: Cell Culture Supernates
  11. Homeostatic chemokines CCL19 and CCL21 promote inflammation in human immunodeficiency virus-infected patients with ongoing viral replication.
    Authors: Damas JK, Landro L, Fevang B, Heggelund L, Tjonnfjord GE, Floisand Y, Halvorsen B, Froland SS, Aukrust P
    Clin. Exp. Immunol., 2009;157(3):400-7.
    Species: Human
    Sample Types: Cell Culture Supernates
  12. CCL19 is constitutively expressed in the CNS, up-regulated in neuroinflammation, active and also inactive multiple sclerosis lesions.
    Authors: Krumbholz M, Theil D, Steinmeyer F, Cepok S, Hemmer B, Hofbauer M, Farina C, Derfuss T, Junker A, Arzberger T, Sinicina I, Hartle C, Newcombe J, Hohlfeld R, Meinl E
    J. Neuroimmunol., 2007;190(1):72-9.
    Species: Human
    Sample Types: Tissue Homogenates
  13. Multiple NF-kappaB and IFN regulatory factor family transcription factors regulate CCL19 gene expression in human monocyte-derived dendritic cells.
    Authors: Pietila TE, Veckman V, Lehtonen A, Lin R, Hiscott J, Julkunen I
    J. Immunol., 2007;178(1):253-61.
    Species: Human
    Sample Types: Cell Culture Supernates
  14. Autologous chemotaxis as a mechanism of tumor cell homing to lymphatics via interstitial flow and autocrine CCR7 signaling.
    Authors: Shields JD, Fleury ME, Yong C, Tomei AA, Randolph GJ, Swartz MA
    Cancer Cell, 2007;11(6):526-38.
    Species: Human
    Sample Types: Cell Culture Supernates
  15. Systemic inflammation in nonalcoholic fatty liver disease is characterized by elevated levels of CCL2.
    Authors: Haukeland JW, Damas JK, Konopski Z, Loberg EM, Haaland T, Goverud I, Torjesen PA, Birkeland K, Bjoro K, Aukrust P
    J. Hepatol., 2006;44(6):1167-74.
    Species: Human
    Sample Types: Serum
  16. Modification of TLR-induced activation of human dendritic cells by type I IFN: synergistic interaction with TLR4 but not TLR3 agonists.
    Authors: Walker J, Tough DF
    Eur. J. Immunol., 2006;36(7):1827-36.
    Species: Human
    Sample Types: Whole Cells
  17. Expression of macrophage inflammatory protein-3beta in human endometrium throughout the menstrual cycle.
    Authors: Daikoku N, Kitaya K, Nakayama T, Fushiki S, Honjo H
    Fertil. Steril., 2004;81(0):876-81.
    Species: Human
    Sample Types: Tissue Homogenates
  18. Expression of macrophage inflammatory protein-3 beta/CCL19 in pulmonary sarcoidosis.
    Authors: Gibejova A, Mrazek F, Subrtova D, Sekerova V, Szotkowska J, Kolek V, du Bois RM, Petrek M
    Am. J. Respir. Crit. Care Med., 2003;167(12):1695-703.
    Species: Human
    Sample Types: BALF
  19. 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

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Human CCL19/MIP-3 beta DuoSet ELISA
By Anonymous on 11/05/2018
Application: Sample Tested: Cell culture supernatant