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Human CXCL16 DuoSet ELISA

Catalog #: DY1164
11 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
DY1164
Ancillary Products Available
DY008B: DuoSet ELISA Ancillary Reagent Kit 2
DY999B: Substrate Reagent Pack
DY999: Substrate Reagent Pack
Human CXCL16 ELISA Standard Curve
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Product Details
Procedure
Citations (11)
FAQs
Supplemental Products
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Summary Product Features Kit Content Other Reagents Required Data Examples Product Datasheets Preparation and Storage Background

Human CXCL16 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Assay Range
15.6 - 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.

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

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


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 equivalent

Reagent Diluent: 1% BSA in PBS, pH 7.2-7.4, 0.2 m filtered (R&D Systems Catalog # DY995). Quality of BSA is critical (see Technical Hints).

Blocking Buffer: 1% BSA in PBS, pH 7.2-7.4, 0.2 m filtered (R&D Systems Catalog # DY995). Quality of BSA is critical (see Technical Hints).

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), or equivalent

Plate Sealers: ELISA Plate Sealers (Catalog # DY992), or equivalent

 

Scientific Data

N/A Human CXCL16 ELISA Standard Curve View Larger

Human CXCL16 ELISA Standard Curve

Product Datasheets

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Product Datasheet
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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: 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.

Entrez Gene IDs:
58191 (Human); 66102 (Mouse)
Alternate Names:
chemokine (C-X-C motif) ligand 16; CXC chemokine ligand 16; CXCL16; CXCLG16; Scavenger receptor for phosphatidylserine and oxidized low density lipoprotein; SCYB16; Small-inducible cytokine B16; SRPSOXC-X-C motif chemokine 16; SR-PSOXTransmembrane chemokine CXCL16

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. Dilute the Capture Antibody (to the working concentration stated in the product datasheet ) 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 each well of the microplate as recommended in the product datasheet. Incubate at room temperature for a minimum of 1 hour.

    Note: The recommended Reagent Diluent typically contains 1% BSA. Some DuoSet Development Kits require alternative blocking agents, or for plates to be blocked overnight with a higher percentage of BSA, please see the product datasheet for details.
     
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

 

PRECAUTION
The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face and clothing protection when using this material.

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 (as recommended in the product datasheet), 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 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.

11 Citations: Showing 1 - 10
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  1. 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: Human
    Sample Types: Cell Culture Supernates
  2. Clinical and biological markers for predicting ARDS and outcome in septic patients
    Authors: J Villar, R Herrán-Mon, E González-H, M Prieto-Gon, A Ambrós, A Rodríguez-, A Muriel-Bom, R Solano, C Cuenca-Rub, A Vidal, C Flores, JM González-M, MI García-Lao, Genetics o
    Scientific Reports, 2021-11-22;11(1):22702.
    Species: Human
    Sample Types: Serum
  3. Intracellular virus sensor MDA5 exacerbates vitiligo by inducing the secretion of chemokines in keratinocytes under virus invasion
    Authors: T Zhuang, X Yi, J Chen, P Kang, X Chen, J Chen, T Cui, Y Chang, Z Ye, Q Ni, Y Wang, P Du, B Li, L Liu, Z Jian, K Li, T Gao, S Li, C Li
    Cell Death Dis, 2020-06-12;11(6):453.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. Differentially Expressed Genes of Natural Killer Cells Can Distinguish Rheumatoid Arthritis Patients from Healthy Controls
    Authors: NM Elemam, MY Hachim, S Hannawi, AA Maghazachi
    Genes (Basel), 2020-04-30;11(5):.
    Species: Human
    Sample Types: Plasma
  5. ADAM-17 is expressed on rheumatoid arthritis fibroblast-like synoviocytes and regulates proinflammatory mediator expression and monocyte adhesion
    Authors: S Ishii, T Isozaki, H Furuya, H Takeuchi, Y Tsubokura, K Inagaki, T Kasama
    Arthritis Res. Ther., 2018-08-02;20(1):159.
    Species: Human
    Sample Types: Serum
  6. Human B cells induce dendritic cell maturation and favour Th2 polarization by inducing OX-40 ligand.
    Authors: Maddur M, Sharma M, Hegde P, Stephen-Victor E, Pulendran B, Kaveri S, Bayry J
    Nat Commun, 2014-06-09;5(0):4092.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. Urine VCAM-1 as a marker of renal pathology activity index in lupus nephritis.
    Authors: Singh S, Wu T
    Oncogene, 2012-07-13;14(4):R164.
    Species: Human
    Sample Types: Urine
  8. Systemic inflammatory markers in COPD: results from the Bergen COPD Cohort Study.
    Authors: Eagan TM, Ueland T, Wagner PD, Hardie JA, Mollnes TE, Damas JK, Aukrust P, Bakke PS
    Eur. Respir. J., 2009-07-30;35(3):540-8.
    Species: Human
    Sample Types: Plasma
  9. Chemokine-enhanced chemotaxis of lymphangioleiomyomatosis cells with mutations in the tumor suppressor TSC2 gene.
    Authors: Pacheco-Rodriguez G, Kumaki F, Steagall WK, Zhang Y, Ikeda Y, Lin JP, Billings EM, Moss J
    J. Immunol., 2009-02-01;182(3):1270-7.
    Species: Human
    Sample Types: BALF
  10. 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: Human
    Sample Types: Urine
  11. Decreased plasma CXCL16/SR-PSOX concentration is associated with coronary artery disease.
    Authors: Sheikine Y, Bang CS, Nilsson L, Samnegard A, Hamsten A, Jonasson L, Eriksson P, Sirsjo A
    Atherosclerosis, 2005-12-27;188(2):462-6.
    Species: Human
    Sample Types: Plasma

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