Abstract：
:Cell-based immunizations are often used when membrane antigens are difficult to purify. To confirm that an antibody binding to the surface of a cell line is, in fact, binding to the desired antigen, FACS can be performed independently on two cell lines, a transfected cell line expressing the antigen of interest and a control cell line not expressing the antigen. Antibodies binding only to the transfected cell line are then selected for further analysis. This approach can be challenging if a large number of antibodies need to be screened and the antibody quantities are limited. Here we describe a novel method that combines the above two steps of FACS screening into a single step, based on the use of two fluorochromes, CMFDA and CMTMR, to stain transfected and control cell lines, respectively. Antibodies conjugated to a thirdfluorochrome are then added to the combined cells, followed by three-color FACS analysis. The newly modified FACS method is simple, sensitive, and high throughput. It can be used for antibody screening in multiple cell lines simultaneously.
journal_name
Biotechniquesjournal_title
BioTechniquesauthors
Yang XP,Gallo M,Ngan I,Nocerini M,Chen MMdoi
10.2144/02323dd05subject
Has Abstractpub_date
2002-03-01 00:00:00pages
678-80, 682issue
3eissn
0736-6205issn
1940-9818journal_volume
32pub_type
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