Mouse CCL21/6Ckine DuoSet ELISA

(12 citations)
(3 Reviews)
  
  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Sample Type & Volume Required
    100 µL
  • Range
    15.60 - 1,000 pg/mL
  • 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.
Ancillary Reagent Kit Available

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant mouse 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.

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)

Preparation and Storage
  • 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);
  • Aliases:
    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, 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:

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.

12 Citations: Showing 1 - 10
Filter your results:

Species
Sample Type
  1. Infection Programs Sustained Lymphoid Stromal Cell Responses and Shapes Lymph Node Remodeling upon Secondary Challenge
    Authors: JL Gregory, A Walter, YO Alexandre, JL Hor, R Liu, JZ Ma, S Devi, N Tokuda, Y Owada, LK Mackay, GK Smyth, WR Heath, SN Mueller
    Cell Rep, 2017;18(2):406-418.
    Species: Mouse
    Sample Type: Tissue Homogenates
  2. Age-Dependent Cell Trafficking Defects in Draining Lymph Nodes Impair Adaptive Immunity and Control of West Nile Virus Infection.
    Authors: Richner J, Gmyrek G, Govero J, Tu Y, van der Windt G, Metcalf T, Haddad E, Textor J, Miller M, Diamond M
    PLoS Pathog, 2015;11(7):e1005027.
    Species: Mouse
    Sample Type: Tissue Homogenates
  3. CCRL1/ACKR4 is expressed in key thymic microenvironments but is dispensable for T lymphopoiesis at steady state in adult mice.
    Authors: Lucas B, White A, Ulvmar M, Nibbs R, Sitnik K, Agace W, Jenkinson W, Anderson G, Rot A
    Eur J Immunol, 2015;45(2):574-83.
    Species: Mouse
    Sample Type: Tissue Homogenates
  4. Plasmodium suppresses expansion of T cell responses to heterologous infections.
    Authors: White C, Villarino N, Sloan S, Ganusov V, Schmidt N
    J Immunol, 2015;194(2):697-708.
    Species: Mouse
    Sample Type: Tissue Homogenates
  5. De novo-induced self-antigen-specific Foxp3+ regulatory T cells impair the accumulation of inflammatory dendritic cells in draining lymph nodes.
    Authors: Alissafi T, Hatzioannou A, Ioannou M, Sparwasser T, Grun J, Grutzkau A, Verginis P
    J Immunol, 2015;194(12):5812-24.
    Species: Mouse
    Sample Type: Tissue Homogenates
  6. The B lymphocyte differentiation factor (BAFF) is expressed in the airways of children with CF and in lungs of mice infected with Pseudomonas aeruginosa.
    Authors: Neill, Daniel R, Saint, Gemma L, Bricio-Moreno, Laura, Fothergill, Joanne L, Southern, Kevin W, Winstanley, Craig, Christmas, Stephen, Slupsky, Joseph R, McNamara, Paul S, Kadioglu, Aras, Flanagan, Brian F
    PLoS ONE, 2014;9(5):e95892.
    Species: Mouse
    Sample Type: Tissue Homogenates
  7. Hypomorphic mutation in the RAG2 gene affects dendritic cell distribution and migration.
    Authors: Maina, Virginia, Marrella, Veronica, Mantero, Stefano, Cassani, Barbara, Fontana, Elena, Anselmo, Achille, Del Prete, Annalisa, Sozzani, Silvano, Vezzoni, Paolo, Poliani, Pietro L, Villa, Anna
    J Leukoc Biol, 2013;94(6):1221-30.
    Species: Mouse
    Sample Type: Tissue Homogenates
  8. Membrane-associated CD93 regulates leukocyte migration and C1q-hemolytic activity during murine peritonitis.
    Authors: Greenlee-Wacker MC, Briseno C, Galvan M, Moriel G, Velazquez P, Bohlson SS
    J. Immunol., 2011;187(6):3353-61.
    Species: Mouse
    Sample Type: Peritoneal Lavage
  9. Induction of targeted cell migration by cutaneous administration of a DNA vector encoding a biologically active chemokine CCL21.
    Authors: Jalili A, Pashenkov M, Kriehuber E, Wagner C, Nakano H, Stingl G, Wagner SN
    J. Invest. Dermatol., 2010;130(6):1611-23.
    Species: Mouse
    Sample Type: Cell Culture Supernates
  10. Role of lymphotoxin-alpha in cigarette smoke-induced inflammation and lymphoid neogenesis.
    Authors: Demoor T, Bracke KR, Maes T, Vandooren B, Elewaut D, Pilette C, Joos GF, Brusselle GG
    Eur. Respir. J., 2009;34(2):405-16.
    Species: Mouse
    Sample Type: BALF
  11. Cyclooxygenase inhibition during allergic sensitization increases STAT6-independent primary and memory Th2 responses.
    Authors: Zhou W, Newcomb DC, Moore ML, Goleniewska K, O'Neal JF, Peebles RS
    J. Immunol., 2008;181(8):5360-7.
    Species: Mouse
    Sample Type: Tissue Homogenates
  12. Therapeutic targeting of CC ligand 21 or CC chemokine receptor 7 abrogates pulmonary fibrosis induced by the adoptive transfer of human pulmonary fibroblasts to immunodeficient mice.
    Authors: Pierce EM, Carpenter K, Jakubzick C, Kunkel SL, Flaherty KR, Martinez FJ, Hogaboam CM
    Am. J. Pathol., 2007;170(4):1152-64.
    Species: Mouse
    Sample Type: Tissue Homogenates
Expand to show all 12 Citations
DuoSet Ancillary Reagent Kits
Description Application Cat# Citations Images  

DuoSet ELISA Ancillary Reagent Kit 2

ELISA DY008 6
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....
Supplemental ELISA Products
Description Application Cat# Citations Images  

Substrate Reagent Pack (8 vials Color A, 8 vials Color B)

ELISA DY999 57
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Reagent Diluent Concentrate 2 (10X, 5 x 21 mL)

ELISA DY995 17
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Stop Solution 2N Sulfuric Acid (15 x 6 mL)

ELISA DY994 12
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

ELISA Plate Sealers, 100 Pack

ELISA DY992
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Streptavidin-HRP (set of 5 vials enough for 75 plates total)

ELISA DY998 44
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Quantikine Wash Buffer 1

ELISA WA126 6
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Clear Polystyrene Microplates, 25 Pack

ELISA DY990 3
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Reagent Additive 1 (21 mL)

ELISA DY005 1
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Glo Substrate Reagent Pack (Reagent A & B)

ELISA DY993 3
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

ELISA Plate-coating Buffer (250 mL of PBS)

ELISA DY006 1
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Reagent Diluent Concentrate 3 (5X, 10 x 21 mL)

ELISA DY004
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....

Black Polystyrene Microplates, 25 Pack

ELISA DY991 2
View Sizes & Prices
Catalog# Size Price Stock Quantity
Loading Information....
Average Rating: 5 (Based on 3 reviews)

Have you used Mouse CCL21/6Ckine DuoSet ELISA?

Submit a review and receive a $25US/€18/£15/$25CAN amazon gift card if you include an image - $10US/€7/£6/$10CAN Amazon card for reviews without an image. Limited to verified customers in USA, Canada and Europe.

Read what people are saying who have used Mouse CCL21/6Ckine DuoSet ELISA.

Filter by Ratings
Excellent
(3)
All Ratings
Filter by Applications
All Applications
Filter by Species
All Species

Images Ratings Applications Species Reviewed By Date Details
  Excellent
  Anonymous 11/19/2016
Show

Summary

Sample TestedCell culture supernatant

Other Experimental Details

Other Experimental DetailsExcellent post-sale service. Many thanks to Drs. Nagarajan and Riedeman, for the excellent technical support.
 Mouse CCL21/6Ckine DuoSet ELISA DY457
Enlarge
Excellent
  Anonymous 05/23/2016
Show
Image Details
 Mouse CCL21/6Ckine DuoSet ELISA DY457
: Mouse CCL21/6Ckine DuoSet ELISA [DY457]

Summary

Sample TestedTissue Homogenates
 Mouse CCL21/6Ckine DuoSet ELISA DY457
Enlarge
Excellent
  Anonymous 05/23/2016
Show
Image Details
 Mouse CCL21/6Ckine DuoSet ELISA DY457
: Mouse CCL21/6Ckine DuoSet ELISA [DY457]

Summary

Sample TestedTissue Homogenates

Order Details

Contact Information

R&D Systems Guarantee

Feedback

Customer Information