The HTRF human CCL20 (MIP3 alpha) kit is designed for the quantification of human CCL20 release in cell supernatant.

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  • No-wash No-wash
  • Low sample consumption Low sample consumption
  • Under 2h bench time Under 2h bench time

The HTRF human CCL20 (MIP3 alpha) kit is designed for the quantification of human CCL20 release in cell supernatant.

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Overview

Also called LARC (liver activation regulated chemokine) or MIP3A (macrophage inflammatory protein 3), CCL20 is a chemokine mainly present in peripheral blood lymphocytes, lymph nodes, and the liver. CCL20 secretion is upregulated by microbial factors and inflammatory cytokines like TNFa or IFNg. Its main role is to attract lymphocytes and dendritic cells to the site of infection. In the CNS, CCL20 released by endothelial cells has been shown to attract TH17 cells and promote their infiltration to the lumbar spinal cord.

Benefits

  • SCALABLE FROM 96 TO 1536WELLS
  • PANEL QUANTIFICATION

Assay principle

Cell supernatant, sample, or standard is dispensed directly into the assay plate for the detection by HTRF® reagents (384-well low-volume white plate or Cisbio low-volume 96-well plate in 20 µl). The antibodies labeled with the HTRF donor and acceptor are pre-mixed and added in a single dispensing step, to further streamline the assay procedure. The assay can be run up toa1536-well format by simply resizing each addition volume proportionally.
CCL20 kit assay principle

Assay data analysis

The 4 Parameter Logistic (4PL) curve is commonly recommended for fitting an ELISA standard curve. This regression enables the accurate measurement of an unknown sample across a wider range of concentrations than linear analysis, making it ideally suited to the analysis of biological systems like cytokine releases.

To fully understand how to deal with HTRF data processing and also 4PL 1/y² fitting, please visit this page.

Cisbio also worked with Myassays.com to help you in your data analysis. By clicking on this link, you’ll be able to access a free online software to run your CCL20 analysis.

Technical specifications of human CCL20 kit

Sample size16 µL
Final assay volume20 µL
Kit componentsLyophilized standard, frozen detection antibodies, buffers &protocol.
LOD &LOQ (in Diluent)15 pg/mL &47 pg/mL
Range47 – 2,000 pg/mL
Time to resultON at RT
SpeciesHuman only

Intra and inter assay

Intra-assay (n=24)

Sample Mean [CCL20] (pg/mL) CV
1 250 8%
2 750 5%
3 1500 4%
Mean CV 6%

Each of the 3 samples was measured 24 times, and % CV was calculated for each sample.

Inter-assay (n=4)

Sample [CCL20] (pg/mL) Mean (delta R) CV
1 263 503 9%
2 593 1124 8%
3 1333 2250 11%
Mean CV 9%

Each of the samples was measured in 4 different experiments, and % CV was calculated for each sample.

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