Human IL1 beta Assay Kit

IL1β, also known as leukocytic pyrogen or mononuclear cell factor, is a pro-inflammatory cytokine of the IL1 family.
IL1β is considered to be a major endogenous pyrogen and induces prostaglandin synthesis, T cell activation, B cell activation and subsequent antibody production. It is also a known promoter of T cell differentiation into Th17.
Cisbio's Human IL1β kit is a homogeneous, add and read assay to measure IL1β 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) 5 pg/mL & 39.4 pg/mL
Range 39.4 – 6,500 pg/mL
Time to result Overnight at RT
Calibration NIBSC (86/680) value (IU/mL) = 0,1 x HTRF hIL1β 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 IL1β analysis.


How to use HTRF cytokine assays

How to measure multiple cytokines from 1 sample

Analytical performance


Intra-assay (n=24)

Sample Mean [IL1β] (pg/mL) CV
1 65 18%
2 705 9%
3 5030 4%
  Mean CV 10%

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

Inter-assay (n=4)

Sample [IL1β] (pg/mL) Mean (delta R) CV
1 159 378 15%
2 702 1742 13%
3 3095 7736 12%
    Mean CV 13%

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


Dilutional linearity

Sample Dilution factor [IL1β] expected (pg/mL) [IL1β] detected (pg/mL) Recovery
1 1 - 5486 -
4.3 1276 1202 94%
10.7 513 479 93%
Mean 93%
2 1 - - -
4.3 408 378 93%
8.9 197 194 99%
18.5 95 95 98%
Mean 97%

The excellent % of recovery obtained from these experiments show the good linearity of the assay.

Spike and recovery

Sample [IL1β] added (pg/mL) [IL1β] expected (pg/mL) [IL1β] detected (pg/mL) Recovery
1 3250 5057 5323 105%
2 3250 7326 7488 102%
      Mean Recovery 104%

The same amount of recombinant cytokine was added to 2 different serum samples, and the set of responses obtained from a standard curve was compared to the calculated expected values. The ~ 100% of recovery observed validates the sample matrix used for this assay.

Assay validation

1. IL1β secretion in THP1 cells stimulated with LPS & PMA: cell densities

THP1 cells plated @ 50 and 100 kcells/well were stimulated for 18h with PMA + LPS respectively used at 50 ng/mL and 1 µg/mL. 16 µL of supernatants were then transferred into a white detection plate to be analyzed by the Human IL1β Assay Kit.

2. IL1β secretion in PBMCs stimulated with LPS: cell density

BPMCs plated @ 50, 100, 200 and 400 kcells/well were stimulated for 18h with various amounts of LPS ranging from 0.02 to 2 µg/mL. 16 µL of supernatants were then transferred into a white detection plate to be analyzed by the Human IL1β Assay Kit.

Part#, inserts & MSDS

Ordering Info

DescriptionCat. noProduct insertMSDS
Human IL1 beta kit - 500 tests62HIL1BPEG
Human IL1 beta kit - 10,000 tests62HIL1BPEH
Human IL1 beta kit - 1 x 96 tests62HIL1BPET

Companion products

DescriptionCat. noProduct insertMSDS
Human IL1 beta kit - 3 Standards62HIL1BCD3
Human IL1 beta kit - Standard62HIL1BCDA