Human CCL21/6Ckine DuoSet ELISA

Catalog # Availability Size / Price Qty
DY366
Ancillary Products Available
Human CCL21 / 6Ckine ELISA Standard Curve
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Product Details
Procedure
Citations (9)
FAQs
Supplemental Products
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Human CCL21/6Ckine 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 CCL21/6Ckine. 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.

Normal Goat Serum: (Catalog # DY005)

 

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)

Normal Goat Serum: (Catalog # DY005)

 

Data Example

Human CCL21 / 6Ckine 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: CCL21/6Ckine

CCL21, also known as 6Ckine, TCA-4, SLC, Exodus-2, and A21, is a homeostatic chemokine that signals through CCR7 or CXCR3. CCL21 is constitutively presented on initial lymphatic vessels, high endothelial venules (HEV), and lymph node dendritic cells (DC) where it promotes the docking of DC to lymphatic vessels and the retention of T cells by lymph node DC, resulting in T cell priming for activation. Its upregulation during chronic inflammation or tissue damage promotes fibrosis, inflammatory cytokine production, and neuropathic pain. The soluble chemokine is elevated in rheumatoid arthritis synovial fluid and in the serum of coronary artery disease patients.

Entrez Gene IDs:
6366 (Human); 18829 (Mouse); 298006 (Rat)
Alternate Names:
6Ckine; 6CkineSmall-inducible cytokine A21; Beta-chemokine exodus-2; CCL21; chemokine (C-C motif) ligand 21; CKb9; exodus-2; member 21; SCYA21; SCYA21MGC34555; secondary lymphoid tissue chemokine; SLC; SLCSecondary lymphoid-tissue chemokine; TCA-4

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 with NGS, 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 CCL21/6Ckine 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.

9 Citations: Showing 1 - 9
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  1. Tissue-engineered 3D melanoma model with blood and lymphatic capillaries for drug development
    Authors: J Bourland, J Fradette, FA Auger
    Sci Rep, 2018;8(1):13191.
    Species: Human
    Sample Types: Cell Culture Supernates
  2. 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
  3. Unbiased Quantitative Proteomics Reveals a Crucial Role of the Allergen Context for the Activation of Human Dendritic Cells
    Authors: L Strasser, HH Dang, H Schwarz, C Asam, F Ferreira, J Horejs-Hoe, CG Huber
    Sci Rep, 2017;7(1):16638.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. 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
  5. 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
  6. 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
  7. 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
  8. Human T-lymphotropic virus type 1-induced CC chemokine ligand 22 maintains a high frequency of functional FoxP3+ regulatory T cells.
    Authors: Toulza F, Nosaka K, Tanaka Y
    J. Immunol., 2010;185(1):183-9.
    Species: Human
    Sample Types: Cell Culture Supernates
  9. Inflammation-induced secretion of CCL21 in lymphatic endothelium is a key regulator of integrin-mediated dendritic cell transmigration.
    Authors: Johnson LA, Jackson DG
    Int. Immunol., 2010;22(10):839-49.
    Species: Human
    Sample Types: Cell Culture Supernates

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