Human CCL1/I-309 DuoSet ELISA Summary
* 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 CCL1/I-309. 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)
Scientific Data
Product Datasheets
Preparation and Storage
Background: CCL1/I-309/TCA-3
I-309, a member of the CC subfamily of chemokines, was initially identified by subtractive hybridization as a transcript that was present in a gamma/delta T cell line, but not in EBV-transformed B cells. Human I-309 is assumed to be a homolog of the mouse TCA-3. While the two proteins share only approximately 42% amino acid sequence identity, both chemokines contain an extra pair of cysteine residues not found in most other chemokines.
Assay Procedure
GENERAL ELISA PROTOCOL
Plate Preparation
- 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.
- 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.
- Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
- Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.
Assay Procedure
- 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.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- 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.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- 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.
- Repeat the aspiration/wash as in step 2.
- Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
- 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 CCL1/I-309 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.
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Citations: Showing 1 - 10
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Dietary Polyphenols Decrease Chemokine Release by Human Primary Astrocytes Responding to Pro-Inflammatory Cytokines
Authors: Grabarczyk, M;Ksiazek-Winiarek, D;Glabinski, A;Szpakowski, P;
Pharmaceutics
Species: Human
Sample Types: Cell Culture Supernates
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Loss of Gasdermin D is protective against influenza virus-induced inflammation and mortality
Authors: S Speaks, A Zani, A Solstad, A Kenney, L Zhang, AC Eddy, AO Amer, R Robinson, C Cai, J Ma, EA Hemann, A Forero, JS Yount
bioRxiv : the preprint server for biology, 2023-04-07;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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Human Primary Astrocytes Differently Respond to Pro- and Anti-Inflammatory Stimuli
Authors: P Szpakowski, D Ksiazek-Wi, M Turniak-Ku, I Pacan, A Glabinski
Biomedicines, 2022-07-22;10(8):.
Species: Human
Sample Types: Cell Culture Supernates
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TSLP regulates mitochondrial ROS-induced mitophagy via histone modification in human monocytes
Authors: YC Lin, YC Lin, ML Tsai, WT Liao, CH Hung
Cell & bioscience, 2022-03-15;12(1):32.
Species: Human
Sample Types: Cell Culture Supernates
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Bacterial genotoxins induce T�cell senescence
Authors: SL Mathiasen, L Gall-Mas, IS Pateras, SDP Theodorou, MRJ Namini, MB Hansen, OCB Martin, CK Vadivel, K Ntostoglou, D Butter, M Givskov, C Geisler, AN Akbar, VG Gorgoulis, T Frisan, N Ødum, T Krejsgaard
Cell Reports, 2021-06-08;35(10):109220.
Species: Human
Sample Types: Cell Culture Supernates
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Immune signatures of protective spleen memory CD8 T cells
Sci Rep, 2016-11-24;6(0):37651.
Species: Human
Sample Types: Cell Culture Supernates
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Activating KIR2DS4 Is Expressed by Uterine NK Cells and Contributes to Successful Pregnancy
Authors: Ashley Moffett
J. Immunol., 2016-11-04;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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Selective class IIa histone deacetylase inhibition via a nonchelating zinc-binding group.
Authors: Lobera, Mercedes, Madauss, Kevin P, Pohlhaus, Denise T, Wright, Quentin, Trocha, Mark, Schmidt, Darby R, Baloglu, Erkan, Trump, Ryan P, Head, Martha S, Hofmann, Glenn A, Murray-Thompson, Monique, Schwartz, Benjamin, Chakravorty, Subhas, Wu, Zining, Mander, Palwinde, Kruidenier, Laurens, Reid, Robert A, Burkhart, William, Turunen, Brandon, Rong, James X, Wagner, Craig, Moyer, Mary B, Wells, Carrow, Hong, Xuan, Moore, John T, Williams, Jon D, Soler, Dulce, Ghosh, Shomir, Nolan, Michael
Nat Chem Biol, 2013-03-24;9(5):319-25.
Species: Human
Sample Types: Cell Culture Supernates
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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., 2011-03-02;186(7):3966-76.
Species: Human
Sample Types: Cell Culture Supernates
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Genetic variants of CC chemokine genes in experimental autoimmune encephalomyelitis, multiple sclerosis and rheumatoid arthritis.
Authors: Ockinger J, Stridh P, Beyeen AD, Lundmark F, Seddighzadeh M, Oturai A, Sorensen PS, Lorentzen AR, Celius EG, Leppa V, Koivisto K, Tienari PJ, Alfredsson L, Padyukov L, Hillert J, Kockum I, Jagodic M, Olsson T
Genes Immun., 2009-10-29;11(2):142-54.
Species: Human
Sample Types: CSF
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FcepsilonRI-mediated thymic stromal lymphopoietin production by interleukin-4-primed human mast cells.
Authors: Okayama Y, Okumura S, Sagara H, Yuki K, Sasaki T, Watanabe N, Fueki M, Sugiyama K, Takeda K, Fukuda T, Saito H, Ra C
Eur. Respir. J., 2009-01-22;34(2):425-35.
Species: Human
Sample Types: Cell Lysates
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Tumor-associated leukemia inhibitory factor and IL-6 skew monocyte differentiation into tumor-associated macrophage-like cells.
Authors: Duluc D, Delneste Y, Tan F, Moles MP, Grimaud L, Lenoir J, Preisser L, Anegon I, Catala L, Ifrah N, Descamps P, Gamelin E, Gascan H, Hebbar M, Jeannin P
Blood, 2007-09-11;110(13):4319-30.
Species: Human
Sample Types: Cell Culture Supernates
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