Human IL10 Assay Kit

Also known as CSIF (Cytokine synthesis inhibitory factor), IL10 is an anti-inflammatory cytokine. Its negative regulation of the immune response is due to its ability to block the signaling of other cytokines, like IFNg, and to inhibit CD28 phosphorylation in T cells.
In immuno-oncology, IL10 is known to inhibit the expansion of TH1 cells while promoting the development of Treg and macrophages.
Cisbio's Human IL10 kit is a homogeneous, add and read assay to measure IL10 secretion in supernatant. It offers a fast, efficient alternative to ELISA.

Sample size 16 µL
Final assay volume 20 µL
Kit components Lyophilized standard, frozen detection antibodies, buffers & protocol.
LOD & LOQ (in Diluent) 7 pg/ml & 22 pg/mL
Range 22 – 6,000 pg/mL
Calibration NIBSC (93/722) value (IU/mL) = 0,005 x HTRF hIL10 value (pg/mL)
Time to result Overnight at RT
Species Human only
Download Insider tips for PBMC isolation Technical Note
Download cytokine release from fresh blood samples application notes

Assay principle

The assay protocol, using a Cisbio 96-well low volume or a 384-well small volume white plate (20 µL final), is described below. Cell supernatant, sample or standard are dispensed directly into the assay plate for the detection by HTRF® reagents. 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 in 96 to 1536-well plates by simply resizing each addition volume proportionally.

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.

Add

Cytokines assay principle add

Read

Cytokines assay principle read

(1) Assay data analysis:

  • "Guidelines for data reduction" button will show you how dealing with HTRF data processing and 4PL 1/y2 fitting.
  • "MyAssays" button will direct you to a free online software able to run hIL10 analysis.

 

How to use HTRF cytokine assays

How to measure multiple cytokines from 1 sample

Analytical performance

Precision

Intra-assay (n=24)

Sample Mean [IL10] (pg/mL) CV
1 62 9%
2 3302 2%
3 5725 3%
  Mean CV 5%

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

Inter-assay (n=4)

Sample [IL10] (pg/mL) Mean (delta R) CV
1 93 717 14%
2 493 4641 11%
3 2609 32182 11%
    Mean CV 12%

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

Part#, inserts & MSDS

Ordering Info

DescriptionCat. noProduct insertMSDS
Human IL10 kit - 500 tests62HIL10PEG
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Human IL10 kit - 10,000 tests62HIL10PEH
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Companion products

DescriptionCat. noProduct insertMSDS
Human IL10 kit - 3 Standards62HIL10CD3
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Human IL10 kit - Standard62HIL10CDA
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