Mouse CXCL16 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 mouse CXCL16. 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: CXCL16
CXC chemokine ligand 16 (CXCL16) is a type I membrane protein that has a non-ELR CXC chemokine domain in its extracellular region. Fractalkine (CX3CL1) and CXCL16 constitute the only two transmembrane chemokines. Mouse and human CXCL16 share 70% amino acid sequence similarity within their chemokine domains and 49% overall amino acid sequence identity.
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 Mouse CXCL16 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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LHPP-mediated inorganic pyrophosphate hydrolysis-driven lysosomal acidification in astrocytes regulates adult neurogenesis
Authors: Sha, L;Li, J;Shen, H;Wang, Q;Meng, P;Zhang, X;Deng, Y;Zhu, W;Xu, Q;
Cell reports
Species: Mouse
Sample Types: Cell Culture Supernates
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Chemokines expressed by engineered bacteria recruit and orchestrate antitumor immunity
Authors: TM Savage, RL Vincent, SS Rae, LH Huang, A Ahn, K Pu, F Li, K de Los San, C Coker, T Danino, N Arpaia
Science Advances, 2023-03-08;9(10):eadc9436.
Species: Mouse
Sample Types: Tissue Homogenates
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Chemotherapy-induced tumor immunogenicity is mediated in part by megakaryocyte-erythroid progenitors
Authors: A Vorontsova, TJ Cooper, J Haj-Shomal, M Benguigui, S Levin, B Manobla, R Menachem, M Timaner, Z Raviv, Y Shaked
Oncogene, 2023-01-16;0(0):.
Species: Mouse
Sample Types: Tissue Homogenates
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Identification of periplakin as a major regulator of lung injury and repair in mice
Authors: V Besnard, R Dagher, T Madjer, A Joannes, M Jaillet, M Kolb, P Bonniaud, LA Murray, MA Sleeman, B Crestani
JCI Insight, 2018-03-08;3(5):.
Species: Mouse
Sample Types: BALF
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NF-?B inducing kinase is a therapeutic target for systemic lupus erythematosus
Authors: HD Brightbill, E Suto, N Blaquiere, N Ramamoorth, S Sujatha-Bh, EB Gogol, GM Castanedo, BT Jackson, YC Kwon, S Haller, J Lesch, K Bents, C Everett, PB Kohli, S Linge, L Christian, K Barrett, A Jaochico, LM Berezhkovs, PW Fan, Z Modrusan, K Veliz, MJ Townsend, J DeVoss, AR Johnson, R Godemann, WP Lee, CD Austin, BS McKenzie, JA Hackney, JJ Crawford, ST Staben, MH Alaoui Ism, LC Wu, N Ghilardi
Nat Commun, 2018-01-12;9(1):179.
Species: Mouse
Sample Types: Serum
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Neutrophils license iNKT cells to regulate self-reactive mouse B cell responses
Nat. Immunol., 2016-10-31;0(0):.
Species: Mouse
Sample Types: Serum
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Lysophosphatidic acid alters the expression profiles of angiogenic factors, cytokines, and chemokines in mouse liver sinusoidal endothelial cells.
Authors: Chou C, Lai S, Ho C, Lin W, Chen C, Lee P, Peng F, Kuo S, Wu S, Lai H
PLoS ONE, 2015-03-30;10(3):e0122060.
Species: Mouse
Sample Types: Cell Culture Supernates
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Elevated urinary VCAM-1, P-selectin, soluble TNF receptor-1, and CXC chemokine ligand 16 in multiple murine lupus strains and human lupus nephritis.
Authors: Wu T, Xie C, Wang HW, Zhou XJ, Schwartz N, Calixto S, Mackay M, Aranow C, Putterman C, Mohan C
J. Immunol., 2007-11-15;179(10):7166-75.
Species: Mouse
Sample Types: Urine
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Acute brain injury triggers MyD88-dependent, TLR2/4-independent inflammatory responses.
Authors: Koedel U, Merbt UM, Schmidt C, Angele B, Popp B, Wagner H, Pfister HW, Kirschning CJ
Am. J. Pathol., 2007-07-01;171(1):200-13.
Species: Mouse
Sample Types: Tissue Homogenates
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Excreted urinary mediators in an animal model of experimental immune nephritis with potential pathogenic significance.
Authors: Wu T, Xie C, Bhaskarabhatla M, Yan M, Leone A, Chen SS, Zhou XJ, Putterman C, Mohan C
Arthritis Rheum., 2007-03-01;56(3):949-59.
Species: Mouse
Sample Types: Urine
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Protein expression pattern in experimental pneumococcal meningitis.
Authors: Klein M, Paul R, Angele B, Popp B, Pfister HW, Koedel U
Microbes Infect., 2006-01-18;8(4):974-83.
Species: Mouse
Sample Types: Tissue Homogenates
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A distinct and unique transcriptional program expressed by tumor-associated macrophages (defective NF-kappaB and enhanced IRF-3/STAT1 activation).
Authors: Biswas SK, Gangi L, Paul S, Schioppa T, Saccani A, Sironi M, Bottazzi B, Doni A, Vincenzo B, Pasqualini F, Vago L, Nebuloni M, Mantovani A, Sica A
Blood, 2005-11-03;107(5):2112-22.
Species: Mouse
Sample Types: Cell Culture Supernates
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