Technical Data
Chloramphenicol Acetyl Transferase (CAT, CatB, CatD, Chloramphenicol Acetyltransferase 2, Chloramphenicol Acetyltransferase II, CAT-II)
Chloramphenicol acetyltransferase (or CAT) is a bacterial enzyme (EC[1] that detoxifies the antibiotic chloramphenicol and is responsible for chloramphenicol resistance in bacteria.[2] This enzyme covalently attaches an acetyl group from acetyl-CoA to chloramphenicol, which prevents chloramphenicol from binding to ribosomes. A histidine residue, located in the C-terminal section of the enzyme, plays a central role in its catalytic mechanism.
The crystal structure of the type III enzyme from Escherichia coli with chloramphenicol bound has been determined. CAT is a trimer of identical subunits (monomer Mr 25,000) and the trimeric structure is stabilised by a number of hydrogen bonds, some of which result in the extension of a beta-sheet across the subunit interface.

Chloramphenicol binds in a deep pocket located at the boundary between adjacent subunits of the trimer, such that the majority of residues forming the binding pocket belong to one subunit while the catalytically essential histidine belongs to the adjacent subunit. His195 is appropriately positioned to act as a general base catalyst in the reaction, and the required tautomeric stabilisation is provided by an unusual interaction with a main-chain carbonyl oxygen.[3]

CAT is used as a reporter system to measure the level of a promoter or its tissue-specific expression. The CAT assay involves monitoring acetylation of radioactively labeled chloramphenicol on a TLC plate; CAT activity is determined by looking for the acetylated forms of chloramphenicol, which have a significantly increased migration rate as compared to the unacetylated form.[4]

Suitable for use in ELISA. Other applications not tested.

Recommended Dilution:
Optimal dilutions to be determined by the researcher.

Storage and Stability:
May be stored at 4°C for short-term only. For long-term storage and to avoid repeated freezing and thawing, add sterile 40-50% glycerol, aliquot and store at -20°C. Aliquots are stable for at least 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
MabIgG2b10B2230Affinity Purified
200ug4°C/-20°C (No Glycerol)Blue IceMouse
Chloramphenicol Acetyl Transferase
Purified by Protein A affinity chromatography.
Supplied as a liquid in PBS, 0.09% sodium azide.
Recognizes chloramphenicol acetyl transferase (CAT), an enzyme involved in bacterial resistance to chloramphenicol.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. Engel J, Prockop DJ (1991). "The zipper-like folding of collagen triple helices and the effects of mutations that disrupt the zipper". Annu. Rev. Biophys. Biophys. Chem. 20 (1): 137–152. doi:10.1146/ PMID 1867713.
2. Shaw WV, Packman LC, Burleigh BD, Dell A, Morris HR, Hartley BS (1979). "Primary structure of a chloramphenicol acetyltransferase specified by R plasmids". Nature 282 (5741): 870–2. doi:10.1038/282870a0. PMID 390404.
3. Leslie AG (1990). "Refined crystal structure of type III chloramphenicol acetyltransferase at 1.75 A resolution". J. Mol. Biol. 213 (1): 167–186. doi:10.1016/S0022-2836(05)80129-9. PMID 2187098.
4. Gorman, CM; Moffat LF; Howard BH (1982). "Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells" (PDF). Mol. Cell. Biol. 2 (9): 1044–1051. doi:10.1128/MCB.2.9.1044. Retrieved 15 October 2012.