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.


Cytokines assay principle add


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


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
Human IL12 kit - 10,000 tests62HIL12PEH

Companion products

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