Introduction of antibody into viable cells using electroporation.
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.
Berglund
D L
DL
Microbiology Department, Montana State University, Bozeman 59717.
Starkey
J R
JR
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000951:Antigens, Neoplasm / Q000276:immunology; D002470:Cell Survival; D005434:Flow Cytometry / Q000379:methods*; D005453:Fluorescence; D008546:Melanoma, Experimental / Q000276:immunology / Q000473:pathology*; D051379:Mice; D015689:Oncogene Protein p21(ras) / Q000276:immunology; D012878:Skin Neoplasms / Q000276:immunology / Q000473:pathology*
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D015689:Oncogene Protein p21(ras)