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Accurate and Simple Measurement of the Pro-inflammatory Cytokine IL-1β using a Whole Blood Stimulation Assay
Published on: March 1, 2011
Measurement of interleukin 6
R P Nordan1, C D Richards, J Gauldie
1National Cancer Institute, Bethesda, Maryland, USA.
Insights
This study presents a sensitive bioassay for quantifying Interleukin 6 (IL-6) across multiple species by measuring cell proliferation. This method offers a viable alternative to ELISA kits for assessing IL-6 bioactivity.
Area of Science:
- Immunology
- Cell Biology
Background:
- Interleukin 6 (IL-6) is a key cytokine involved in immune responses and cellular processes.
- IL-6 is produced by various cells upon stimulation, highlighting its broad biological significance.
Purpose of the Study:
- To describe a simple and sensitive bioassay for quantifying Interleukin 6 (IL-6) in multiple species.
- To provide protocols for maintaining cell lines and preparing reagents for IL-6 measurement.
Main Methods:
- Utilized an IL-6-dependent murine B cell hybridoma cell line (B9) to assess IL-6 bioactivity.
- Developed protocols for B9 cell maintenance, IL-6 standard preparation, and supernatant collection.
Main Results:
- Established a sensitive assay for human, rat, rabbit, pig, and mouse IL-6.
- Demonstrated the utility of the B9 cell proliferation assay for IL-6 quantification.
Conclusions:
- The described bioassay offers a sensitive and species-versatile method for measuring IL-6 bioactivity.
- ELISA kits are an alternative for human/mouse IL-6 measurement but do not assess bioactivity.
Abstract:
Interleukin 6 (IL-6) is a pluripotent cytokine with multiple effects on many different cell types. It is produced by a variety of cells in response to immunological and other stimuli. This unit describes a simple and sensitive assay for human, rat, rabbit, pig, and mouse IL-6, based on IL-6-dependent proliferation of a murine B cell hybridoma cell line, B9. Support protocols discuss maintenance of B9 cells, preparation of IL-6 standards, and production of IL-6-containing supernatant. In addition, IL-6 ELISA kits for the measurement of human or mouse IL-6 are available from a number of companies. These products are reliable, highly sensitive, and specific, and thus should be considered as an excellent (although more expensive) alternative, keeping in mind that bioactivity is not assessed with this approach.
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