Application of NF-?B reporter and Dual-Luciferase assays in the measure of bioactivity of interleukin-1? and interleukin-1 receptor antagonist
- VernacularTitle:利用NF-?B转录活性萤光素酶报告系统检测白细胞介素1及白细胞介素1受体拮抗剂的生物活性
- Author:
Yingmei ZHANG
;
Ying WANG
;
Qianmei XU
;
Bingfeng LV
- Publication Type:Journal Article
- Keywords:
NF-kappaB;
Interleukin-1;
Receptors,interleukin-1;
Luciferase
- From:
Journal of Peking University(Health Sciences)
2003;0(06):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To develop a reporter gene system based on transient transfections with a NF-?B responsive reporter gene to detect the bioactivity of IL-1? and IL-1 receptor antagonist.Methods: NF-?B reporter and Dual-Luciferase assays were applied to measure the bioactivity of IL-1? and IL-1 receptor antagonist in mouse EL4 cells(some subclones of EL4 cells expressed high level of IL-1 receptor on cell surface).pNF-?B-luc and pRL-TK, used as an internal control,were co-transfected into EL4 cells and then the IL-1? was added.Results: The results indicated that IL-1? was able to induce the expression of this luciferase,which could be blocked by IL-1 receptor antagonist. The optimal dose of IL-1? was 5(?g/L) in Dual-Luciferase assay,whose bioactivity can be effectively inhibited by IL-1ra at 50 ?g/L.Conclusion: We have established a new method to detect the bioactivity of IL-1? and IL-1 receptor antagonist,which can give repeatable results.
