Human IL1 beta kit HTRF®

The HTRF human IL beta kit is designed for the quantification of human IL1 beta release in cell supernatant.

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  • No-wash No-wash
  • Low sample consumption Low sample consumption
  • Under 2h bench time Under 2h bench time

The HTRF human IL beta kit is designed for the quantification of human IL1 beta release in cell supernatant.

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IL1 beta, also known as leukocytic pyrogen or mononuclear cell factor, is a pro-inflammatory cytokine of the IL1 family. IL1 beta 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.

Assessment of serum samples often requires enhanced sensitivity. In some cases, AlphaLISA assays may have sufficient sensitivity to enable the detection of low levels of analytes in serum or plasma. Check out the   Human IL-1β AlphaLISA kit’s analytical performances for more information or learn more about AlphaLISA. When assaying, always follow the recommended protocol and avoid highly haemolyzed samples.



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.
IL1 beta kit assay principle

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 IL1 beta analysis.

Technical specifications of human IL1 beta kit

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 hIL1ß value (pg/mL)
Species Human only

Intra and inter assay

Intra-assay (n=24)

Sample Mean [IL1ß] (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 [IL1ß] (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

Dilution factor[IL1β] expected (pg/mL)[IL1β] detected (pg/mL)Recovery

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

Spike and recovery

[IL1β] added (pg/mL)[IL1β] expected (pg/mL)[IL1β] detected (pg/mL)Recovery

Mean Recovery104%

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.

IL1ß secretion in THP1 cells stimulated with LPS & PMA

THP1 cells plated at 50 and 100 kcells/well were stimulated for 18 h with PMA and LPS respectively used at 50 ng/mL and 1 µg/mL. 16 µL of supernatants were then transferred into a white detection plate (384 low volume) to be analyzed with the Human IL1ß Assay.

IL1ß secretion in THP1 cells stimulated with LPS and PMA

IL1ß secretion in PBMC stimulated with LPS

PBMC plated at 50, 100, 200 and 400 kcells/well were stimulated for 18 h with increasing concentrations of LPS ranging from 0.02 to 2 µg/mL. 16 µL of supernatants were then transferred into a white detection plate (384 low volume) to be analyzed by the Human IL1ß Assay Kit.

IL1ß secretion in PBMC stimulated with LPS

Celular High Throughput Screening assays using HTRF detection of cytokines

In collaboration with GSK - Présentations scientifiaues

HTRF Cytokine cell-based assays

Monitor immune regulation the simple way - Flyers

HTRF assays for Oncology and Inflammation

Signaling in the immune system - Brochures

HTRF measurement of cytokine release from fresh blood samples

In collaboration with Blood assay solution - Notes d'application

Cytokine secretion profile on PBMCs

One sample to measure them all - Posters

Guidelines from PBMC isolation to cytokine assay optimisation

Perform and optimize cytokine assays on PBMC - Notes techniques

How to freeze PBMC

Validated protocol - Notes techniques

Webinar: NASH - Fibrosis Research and Drug Discovery

Featuring a panel of experts - Vidéos

How to measure multiple cytokines from 1 sample

What to expect at the bench - Vidéos

HTRF makes cytokine detection easy

A fun video introducing you to cytokines assays with HTRF - Vidéos

STING HTRF offer to bridge innate and adaptive immunity

cGAS-STING signaling pathway from A to Z - Brochures

Novel HTRF platform to delineate STING pathway

Explore the whole STING pathway with a single technology - Posters

Cisbio Product Catalog 2019

All your HTRF assays in one document! - Catalogue

A guide to Homogeneous Time Resolved Fluorescence

General principles of HTRF - Guides

Cytokine assays: a guide to success with HTRF

Setting up a successful cytokine assay - Guides

Functional response to PD1 blockade rapidly assessed through HTRF IFNγ quantification

For researcher working on checkpoint inhibitors - Notes d'application

Portfolio of HTRF cytokine assays

This leaflet presents the analytical performances of each assay - Brochures

How HTRF compares to Western Blot and ELISA

Get the brochure about technology comparison. - Brochures

Protocol for Th17 differentiation from CD4 mouse T-lymphocyte

A step by step protocol for Th17 cells differentiation from naive CD4 mouse cells - Notes techniques

An innate and adaptive immunity recap

Insight into the diversity of cells & signaling pathways - Guides

Molecular basis of neuroinflammation and neurodegeneration diseases

The essential guide for extending your knowledge on the molecular mechanisms of neurodegenerative diseases - Guides

Note about HMGB1 assay

HTRF HMGB1, IL-1β and IL-18 assays discriminate between inflammasome triggered pyroptosis, and necroptosis - Notes d'application

On-demand webinar: Linking Neuroinflammation and Neurodegeneration

New insight into neuroinflammation research - Vidéos

Assays for autoimmunity research

Advance your research on autoimmune diseases - Flyers

Plate Reader Requirement

Choosing the right microplate reader ensures you’ll get an optimal readout. Discover our high performance reader, or verify if your lab equipment is going to be compatible with this detection technology.

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