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

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Overview

Mainly produced by leukocytes, IFN alpha is involved in innate immunity response to viral infection. IFN alpha activates several intracellular pathways in target cells, which explains its pleiotropic effects on cell differentiation, growth, and immunoregulation.


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 IFN-α 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.

Benefits

  • SCALABLE FROM 96 TO 1536WELLS
  • PANEL QUANTIFICATION

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.
IFN alpha 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 IFN alpha analysis.

Technical specifications of human IFN alpha kit

Sample size 16 µL
Final assay volume 20 µL
Kit components Lyophilized standard, frozen detection antibodies, buffers &protocol.
LOD &LOQ (in Diluent) 11 pg/mL &36 pg/mL
Calibration NIBSC (94/784) value (IU/mL) = 0,9 x HTRF hIFNa HTRF
Range 36 – 4,000 pg/mL
Time to result 2h at RT
Species Human only

Intra and inter assay

Intra-assay (n=24)

SampleMean [IFNa] (pg/mL)CV
115011%
215002%
330005%

Mean CV6%

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

Inter-assay (n=4)

Sample [IFNa] (pg/mL) Mean (delta R) CV
1 47 268 9%
2 206 1150 5%
3 1905 4227 4%
Mean CV 6%

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

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In collaboration with Blood assay solution - Application Notes

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 - Technical Notes

How to freeze PBMC

Guideline for PBMC freezing - Technical Notes

How to measure multiple cytokines from 1 sample

What to expect at the bench - Videos

Product Insert IFN a (h) Kit / 62HIFNAPEG-62HIFNAPEH

62HIFNAPEG-62HIFNAPEH - Product Insert

HTRF makes cytokine detection easy

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

Cisbio Product Catalog 2019

All your HTRF assays in one document! - Catalog

A guide to Homogeneous Time Resolved Fluorescence

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Cytokine assays: a guide to success with HTRF

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This leaflet presents the analytical performances of each assay - Brochures

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Get the brochure about technology comparison. - Brochures

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Safety Data Sheet IFN a (h) Kit / 62HIFNAPEG

62HIFNAPEG - Safety Data Sheet

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