Berglund D L, Starkey J R
Microbiology Department, Montana State University, Bozeman 59717.
Cytometry. 1991;12(1):64-7. doi: 10.1002/cyto.990120109.
Conditions for labelling an intracellular antigen, p21ras, using electroporation to introduce a fluorescent antibody, are described. Following labelling, cells were evaluated for p21ras associated fluorescence by flow cytometry. Electroporation, sorting, and cell handling parameters were varied to determine optimal conditions for cell viability. Cells were best held in serum containing growth medium both before and after electroporation, while antibody introduction during the electroporation phase was most efficient when carried out in a balanced saline solution. For maximum efficiency of antibody internalization, the antibody needed to be present during electroporation, and medium needed to be replaced several times in the first few hours after electroporation to ensure good cell survival.
本文描述了使用电穿孔法引入荧光抗体标记细胞内抗原p21ras的条件。标记后,通过流式细胞术评估细胞中与p21ras相关的荧光。改变电穿孔、分选和细胞处理参数以确定细胞活力的最佳条件。电穿孔前后,细胞最好保存在含血清的生长培养基中,而在电穿孔阶段于平衡盐溶液中引入抗体时效率最高。为了使抗体内化效率最大化,在电穿孔期间需要存在抗体,并且在电穿孔后的最初几个小时内需要更换几次培养基以确保细胞良好存活。