摘要
目的用异硫氰酸荧光素(FITC)标记抗马干扰素-γ(IFN-γ)单克隆抗体,为建立马IFN-γ细胞内细胞因子染色方法提供实验材料。方法利用FITC标记纯化的抗马IFN-γ单克隆抗体(SB10),荧光抗体经系列稀释后,对马外周血单核细胞(PBMC)进行细胞内细胞因子IFN-γ单染色的流式细胞术检测,同时与商品化FITC标记的抗牛IFN-γ单克隆抗体(CC302)进行流式细胞术检测对比试验。结果FITC-CC302染色马PBMC,1μl荧光抗体染色106个细胞时,分泌IFN-γ的细胞频率为9.35%,而FITC-SB10经系列稀释染色,在用0.25μl荧光抗体染色106个细胞时,分泌IFN-γ的细胞频率为9.90%。结论所制备的FITC标记的抗马IFN-γ单克隆抗体,可为建立马细胞免疫学研究方法奠定物质基础,进而为建立监测马机体免疫状态和研究机体免疫机制提供技术平台。
Objective To label equine IFN-γ monoclonal antibody（McAb） with fluorescein isothioeyanate（FITC） and provide an experimental material for intracellular eytokine staining （ICS） assay of IFN-γ in equine lymphocytes. Methods Purified equine IFN-γ MeAb SB10 was labeled with FITC, then diluted serially and used for ICS assay of IFN-γ in equine PBMCs by flow eytometry. The result was compared with that of flow cytometry by using commercial FITC-labeled bovine IFN-γ MeAb CC302. Results When 106 PBMCs were stained with 1 μl of FITC-CC302, the frequency of PBMCs secreting IFN-γwas 9. 35%. However, when 106 PBMCs were stained with 0. 25 μl of FITC-SB10, the frequency was 9. 90%. Conclusion The prepared FITC-labeled equine IFN-γ McAb laid a material foundation of developing the methods for immunological study on equine cells and provided a technical platform for monitoring the immune state and investigating the immunological mechanism.
出处
《中国生物制品学杂志》
CAS
CSCD
2009年第2期188-191,共4页
Chinese Journal of Biologicals
基金
国家"863"计划(2006AA10A203)
黑龙江省自然科学基金重点项目(ZJN-0602-01)
关键词
马干扰素-γ
单克隆抗体
异疏氰酸荧光素
细胞内细胞因子染色
Equine IFN-γ
Monoclonal antibody
Fluorescein isothiocyanate （FITC）
Intracellular eytokine staining （ICS）
