胞内细胞因子最佳染色抗体浓度的选择

Selecting the Optimal Concentration of Immunofluorescent Antibodies for Intracellular Cytokine Staining

探讨胞内细胞因子最佳染色浓度的选择方法。以人外周血单个核细胞为研究对象 ,应用胞内免疫荧光染色和流式细胞仪检测分析 ,比较不同浓度商品化IFNγ -FITC单抗和IL4 -FITC单抗与实验室制备的IFNγ -FITC单抗和IL4 -FITC单抗的荧光染色效果。结果表明 ,应用不同浓度商品化抗体得到的染色率不同。选择与用商品化抗体最佳染色浓度得到的染色率相同的实验室制备荧光抗体的染色浓度作为实验室制备荧光抗体的最佳染色浓度。

The aim of this study was to find an optimal concentration of immunofluorescent antibodies for intracellular cytokine staining.The expression of intracellular IFNγ and IL4 in normal human peripheral blood mononuclear cells was detected by intracellular staining and flow cytometry(FCM).The result of flow cytometry was indicated that positive staining rates obtained by applying commercial immunofluorescent antibodies to IFNγ and IL4 were significantly different from that by applying our lab-made immunofluorescent antibodies to IFNγ and IL4.Moreover, with the increase in the antibody concentration, the positive rates varied.Therefore, we conclude that the optimal concentration of lab-made immunofluorescent antibodies for intracellular cytokine should be the one that can obtain the same positive staining rates as that obtained by using their commercial counterparts.