A microtiter assay for determining protein, acetylcholinesterase activity, and G418 (Neomycin) resistance in cultured cells.
The Coomassie brilliant blue assay for the determination of protein has been extended to rapidly and conveniently measure the protein concentration of cells growing in culture in a 96-well microtiter format. Modifications of the standard assay include sodium hydroxide to solubilize the cells and ovalbumin, instead of bovine serum albumin, as a protein standard. The procedure allows a large number of small samples to be assayed simultaneously. Two examples of its use, enzyme-specific activity and drug resistance, are shown. An assay for acetylcholinesterase activity in the same culture plate is demonstrated. G418, an inhibitor of cell protein synthesis, is frequently used to select for cells transfected with the neomycin resistance gene. The required concentration of G418 can be easily determined with this protein assay.
Elson
H F
HF
Division of Biochemistry, Walter Reed Army Institute of Research, Washington, D.C. 20307.
Gentry
M K
MK
Doctor
B P
BP
D000110:Acetylcholinesterase / Q000032:analysis*; D000818:Animals; D002455:Cell Division; D002478:Cells, Cultured; D002642:Chick Embryo; D003124:Colorimetry / Q000379:methods; D004352:Drug Resistance, Microbial; D008832:Microchemistry; D009132:Muscles / Q000166:cytology / Q000378:metabolism; D009355:Neomycin; D011506:Proteins / Q000032:analysis*; D012394:Rosaniline Dyes; D013194:Staining and Labeling; D014162:Transfection
D011506:Proteins; D012394:Rosaniline Dyes; D000110:Acetylcholinesterase; D009355:Neomycin; C004692:coomassie Brilliant Blue