Rapid, sensitive, and inexpensive assay for chloramphenicol acetyltransferase

Journal Article · Wed Apr 01 04:00:00 EST 1987 · DNA; (United States)

DOI:https://doi.org/10.1089/dna.1987.6.173· OSTI ID:6477239

Nordeen, S K; Green, III, P P; Fowlkes, D M

We present a rapid, sensitive enzymatic assay for chloramphenicol acetyltransferase (CAT) that does not require chromatography, HPLC, or autoradiography. The assay is based on the use of an inexpensive substrate, tritiated acetate, instead of (/sup 14/C)chloramphenicol. The method is adapted from one originally used by de Crombrugghe et al. and by Shaw, but with simplifications appropriate for routine use. In our hands, the method is as sensitive as the customary thin-layer chromatography assay and is far more efficient for the performance of many assays, both in terms of labor and expense.

• Research Organization: Univ. of Colorado Health Sciences Center, Denver
• OSTI ID: 6477239
• Journal Information: DNA; (United States), Journal Name: DNA; (United States) Vol. 2; ISSN DNAAD
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ACETATES
ANIMALS
CARBON 14 COMPOUNDS
CARBOXYLIC ACID SALTS
CHROMATOGRAPHY
ENZYMES
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
LIQUID COLUMN CHROMATOGRAPHY
MAMMALS
MICE
RADIOENZYMATIC ASSAY
RODENTS
SENSITIVITY ANALYSIS
SEPARATION PROCESSES
TRACER TECHNIQUES
TRANSFERASES
TRITIUM COMPOUNDS
VERTEBRATES