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Detection and Isolation of Viable Mouse IL-17-Secreting T Cells
Published on: December 18, 2008
Sandwich enzyme immunoassay for murine IL-3
H J Ziltener1, I Clark-Lewis, S L McDonald
1Biomedical Research Centre, UBC, Vancouver, Canada.
Cytokine
|November 1, 1989
Summary
A new immunoassay accurately detects murine interleukin-3 (IL-3) in just 6 hours. This sensitive method offers a faster alternative to traditional bioassays for quantifying IL-3 levels.
Area of Science:
- Immunology
- Biochemistry
Background:
- Murine interleukin-3 (IL-3) is a critical cytokine for myeloid cell development.
- Existing bioassays for IL-3 are time-consuming, requiring 2-3 days for results.
Purpose of the Study:
- To develop a reproducible and sensitive immunoassay for quantifying murine IL-3.
- To provide a faster alternative to current IL-3 detection methods.
Main Methods:
- Development of a sandwich immunoassay utilizing two polyclonal antipeptide antibody preparations.
- Antibodies were raised against synthetic N-terminal (1-29) and full-length (1-140) IL-3 peptides.
- Assay optimization for sensitivity and reproducibility across various IL-3 sources.
Main Results:
- The immunoassay demonstrated high sensitivity, detecting as little as 2-4 U/ml of bioactive IL-3 or approximately 60 pg/ml of synthetic IL-3.
- The assay reliably detected IL-3 from diverse sources, including T cells, WEHI-3B cells, and recombinant sources.
- Assay completion time was significantly reduced to 5-6 hours, compared to days for bioassays.
Conclusions:
- A novel, rapid, and sensitive immunoassay for murine IL-3 has been successfully developed.
- This assay offers a significant improvement in speed and efficiency for IL-3 quantification.
- The developed immunoassay is suitable for detecting IL-3 from various biological and recombinant sources.
