Cold Spring Harb Protoc. 2021 Jun 1;2021(6):2021/6/pdb.prot099697. doi: 10.1101/pdb.prot099697.
Staining of suspension cells after fixation, described here, is normally used only to detect cell-surface antigens. Once cells are fixed and permeabilized, the antibodies are added. The antibodies can be labeled directly or they can be detected by using a labeled secondary reagent that will bind specifically to the primary antibody. Detection reagents for cell staining can be labeled with fluorochromes, enzymes, gold, or iodine. Because the cells are in suspension, the washing steps are tedious, and care should be taken not to centrifuge for long durations or at high speeds.
这里描述的固定后悬浮细胞染色通常仅用于检测细胞表面抗原。一旦细胞固定和通透，就添加抗体。抗体可以直接标记，也可以通过使用与初级抗体特异性结合的标记的二级试剂来检测。细胞染色的检测试剂可以用荧光染料、酶、金或碘标记。由于细胞处于悬浮状态，洗涤步骤很繁琐，应注意不要长时间或高速离心。
