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HTRF Human IFNβ Detection Kit, 500 Assay Points

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

For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption & disposal requirements under European REACH regulations (EC 1907/2006).

Product Variants
Part number: 62HIFNBPEG
Unit Size: 500 Assay Points
List price: USD 1,370.00
Your price:
USD 1,370.00
USD 1,370.00 /each
Part number: 62HIFNBPEH
Unit Size: 10,000 Assay Points
List price: USD 14,590.00
Your price:
USD 0.00
USD 14,590.00 /each

Overview

Type I interferons, interferon alpha (IFN-α) and interferon beta (IFN-β), play essential roles in innate immune response to viral infections. IFN-β is produced by fibroblasts, macrophages, and dendritic cells, as well as endolial cells. expression of IFN-β gene is controlled by transcription factors like IRF-3, c-Jun/ATF-2, and NF-kappaB downstream of Pattern Recognition Receptors, such as Toll-like receptors. In particular, IFN-β is primary functional readout of cGas-cGAMP-STING pathway activation. Once secreted, IFN-β binds and activates IFNAR1/IFNAR2 receptor complex, which relies on STAT1/2 transcription factors. Phosphorylated STAT1/2 leads to induction of anti-viral gene expression.

Assessment of serum samples often requires enhanced sensitivity. In some cases, AlphaLISA assays may have sufficient sensitivity to enable detection of low levels of analytes in serum or plasma.

Specifications

Assay Points
500
Assay Target Type
Kit
Assay Technology
HTRF
Brand
HTRF
Quantity
1
Shipping Conditions
Shipped in Dry Ice
Therapeutic Area
Oncology & Inflammation
Unit Size
500 Assay Points

Video gallery

How it works

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 Revvity 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.

ifn beta
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.

Assay details

Technical specifications of human IFN beta kit
Sample size 14 µL
Final assay volume 20 µL
Kit components Lyophilized standard, frozen detection antibodies, activation reagent, buffers & protocol
LOD & LOQ (in Diluent) 7 pg/mL & 19 pg/mL
Range 19 – 4,000 pg/mL
Time to result 3 h at RT
Calibration NIBSC (00/572) value (IU/ml) = 0.44 x HTRF hIFNβ value (pg/ml)
Species Human only

 

Analytical performance

Intra and inter assay

Intra assay (n=24)

Sample Mean [IFN-β] (pg/mL) CV
1 33 4%
2 321 3%
3 3900 5%
  Mean CV 4%

 

Inter assay (n=4)

Sample [IFN-β] (pg/mL) Mean (delta ratio) CV
1 62 374 10%
2 329 1628 7%
3 1739 5690 9%
    Mean CV 9%

 

Assay validation

STING pathway activation by cGAMP triggers hIFNβ release in PBMC

400,000 PBMC were seeded in a 96 well plate under 100 µL of cell culture medium, then stimulated with 50 µL of cGAMP used at 67µM. After an overnight incubation, 14 µL of supernatant were transferred into a white detection plate and human IFNβ was quantified by HTRF.

assay validation - human ifn beta 1

 

​LPS efficiently induces hIFNβ release in THP1 cells

400,000 THP1cells were seeded in a 96 well plate under 100 µL, then stimulated with 50 µL of LPS used at 20 µg/ml. After 4h30 incubation, 14 µL of supernatant were transferred into a white detection plate, and human IFNβ was quantified by HTRF.

human ifn beta 2

 

Recombinant THP1 cells expressing constitutively activated STING variant spontaneously releases hIFNβ

400,000 of THP1-WT cells (wild type) and THP1-S154 cells (THP1-Dual™ KI-hSTING-S154 Cells, Invivogen) were seeded in a 96 well plate, under 100 µL of cell culture medium. After 4 h 30 of stimulation, 14 µl of supernatant were transferred into a white detection plate, and human IFNβ was quantified by HTRF.

assay validation - human ifn beta 3

 

dsDNA mimetic (CDS agonist) induces hIFNβ secretion

400,000 THP1-S154 cells (THP1-Dual™ KI-hSTING-S154 Cells, Invivogen) were seeded in a 96 well plate under 100 µL of cell culture medium, then treated with dsDNA mimetic (CDS Agonist – LyoVec, Invivogen). After an overnight incubation, 14 µL of supernatant were transferred into a white detection plate, and human IFNβ was quantified by HTRF.

assay validation - human ifn beta 4

Resources

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Application Note Icon
Application Note
Download your application note on STING agonists characterization with IFN-β

Discover a rapid and reliable tool for the identification and characterization of STING-targeting compounds.

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