Detection of chloramphenicol acetyl transferase activity in transfected cells: a rapid and sensitive HPLC-based method.
An assay based on high-performance liquid chromatography (HPLC) has been developed for determining the expression of the acetyl coenzyme A: chloramphenicol 3-O-acetyltransferase gene in transfected cells. The assay is based on the separation of extracted, nonradioactive chloramphenicol and its acetylated derivatives by reverse-phase HPLC, using UV absorbance for detection. In addition to eliminating the need for costly [14C]chloramphenicol, this assay is as sensitive and much faster than the commonly used method involving thin-layer chromatography.
D000123:Acetyltransferases / Q000032:analysis / Q000235:genetics*; D000818:Animals; D002478:Cells, Cultured; D015500:Chloramphenicol O-Acetyltransferase; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002855:Chromatography, Thin Layer; D005819:Genetic Markers; D014158:Transcription, Genetic; D014162:Transfection
D005819:Genetic Markers; D000123:Acetyltransferases; D015500:Chloramphenicol O-Acetyltransferase