Antigens expressed on live cells

Soluble recombinant protein extracted fiom cells or supernatants of eukaryotic cells (e.g, Chinese hamster ovary cells or insea cells expressing recombinant baculovlrus) or baaeria (such as . colt harbouring plasmids or recombinant viruses) and dissolved in PBS Protein separated electrophoretically in sodium dodecyl sulfate-containing polyacrylamide gels (SDS-PAGE) and eluted from gel slices into PBS Cultured cells grown in suspension or as monolayers in flasks, then detached by treatment with 0,1% trypsin-EDTA, and resuspended in PBS or serum-free DMEM 

Freund s complete adjuvant (FCA) Freund s incomplete adjuvant (FIA) 

Mix proteins, peptide conjugates, or eluates from polyacrylamide gels in PBS 1 1 with adjuvant (FCA for the first immunization. FIA for subsequent immunizations) in a capped plastic tube (LP3, Bijou, or 30 ml Universal) by vortexing until a stable emulsion is formed. 

Check that phase separation does not occur on standing at 4 C for 1 h. Alternatively, allow a drop of emulsion to fall from a Pasteur pipette onto a water surface the drop should contract, remain as a droplet, and not disperse. 

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