Human IL12 Assay Kit

Also known as NKSF (natural killer cell stimulatory factor) and CLMF (cytotoxic lymphocyte maturation factor), IL12 is produced by macrophages and B cells.
Along with IFNg, IL12 promotes the differentiation of naïve T cells into TH1 cells, which in turn release IFNg and IL2. Because of its effects on TH1 induction, IL12 is strongly linked to autoimmunity.
IL12 is a heterodimer of p35 and p40 which is shared with IL23. Cisbio's IL12 assay is specific for IL12p70 (full form) and does not cross react with IL23
The kit is a homogeneous, add and read assay to measure IL12 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) 14 pg/mL & 81 pg/mL
Range 55 – 13,000 pg/mL
Time to result 2h at RT
Calibration NIBSC (95/544) value (IU/mL) = 0,01 x HTRF hIL12 value (pg/mL)
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.

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Cytokines assay principle add

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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 hIL12 analysis.

 

How to use HTRF cytokine assays

How to measure multiple cytokines from 1 sample

Analytical performance

Precision

Intra-assay (n=24)

Sample Mean [IL12] (pg/mL) CV
1 200 6%
2 5000 4%
3 10000 3%
  Mean CV 4%

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

Inter-assay (n=4)

Sample [IL12] (pg/mL) Mean (delta R) CV
1 252 444 8%
2 1221 2158 11%
3 5909 10789 12%
    Mean CV 10%

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 IL12 kit - 500 tests62HIL12PEG
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Human IL12 kit - 10,000 tests62HIL12PEH
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Companion products

DescriptionCat. noProduct insertMSDS
Human IL12 kit - 3 Standards62HIL12CD3
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Human IL12 kit - Standard62HIL12CDA
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