Kupper H, Storz H
Acta Histochem. 1986;78(2):185-8. doi: 10.1016/S0065-1281(86)80052-6.
A simple method for double staining by immunofluorescence is described. If for double staining using monoclonal antibodies of the same species only one antibody is conjugated with FITC or TRITC, a combination of indirect and direct immunofluorescence is possible. For cell staining the following incubation steps are carried out: Monoclonal antibody I (unlabelled, mouse), anti-mouse immunoglobulin serum FITC- or TRITC-conjugated, normal mouse serum for blocking of free binding sites of the anti-mouse immunoglobulin, and monoclonal antibody II (mouse) which is conjugated with an alternative fluorochrome. The use of this method is demonstrated for investigation of single cell suspensions (performed as a slide test) and of cryostat sections.
本文描述了一种简单的免疫荧光双重染色方法。如果使用同一种属的单克隆抗体进行双重染色，仅将一种抗体与异硫氰酸荧光素（FITC）或四甲基异硫氰酸罗丹明（TRITC）偶联，则间接免疫荧光和直接免疫荧光相结合是可行的。对于细胞染色，进行以下孵育步骤：单克隆抗体I（未标记，小鼠来源）、异硫氰酸荧光素或四甲基异硫氰酸罗丹明偶联的抗小鼠免疫球蛋白血清、用于封闭抗小鼠免疫球蛋白游离结合位点的正常小鼠血清，以及与另一种荧光染料偶联的单克隆抗体II（小鼠来源）。本文展示了该方法在单细胞悬液（作为载玻片试验进行）和冰冻切片研究中的应用。
