The Human IL12 kit is designed for the quantification of IL12 release in cell supernatant.
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  • No-wash
  • Low sample consumption
  • Highly specific
The Human IL12 kit is designed for the quantification of IL12 release in cell supernatant.
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In stock

Overview

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 IFN gamma, 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 p4,0 which is shared with IL23. Cisbio's IL12 assay is specific for IL12p70 (full form) and does not cross react with IL23.

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 to a 1536-well format by simply resizing each addition volume proportionally.

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 IL12 analysis.

Technical specifications of human IL12 kit

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 81 – 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

Intra and inter assay

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.

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