Skatole → 4-(1H-Indol-2-yl)butanoic acid

Participants

Reaction Steps

Microtiter plates (Nunc, Sweden) coated with anti-indoleacetic acid polyclonal antibodies (described above) were used. A mixture of 100 μl of the enzyme conjugate (Indole-3-butyric acid-AP conjugate, prepared as described above, and diluted 1:10,000 in TBS) and 100 μl of skatole solution (either in TBS or in 10% porcine serum solution) of a number of dilutions was added to wells in duplicate. The mixture was incubated for 3 hrs. at 4° C. and then washed with saline-Tween solution 3 times (200 μl volume). 200 μl of the substrate solution (1mg/ml in TBS) was added per well and incubated at 37° C. for an hour. The wells were then treated with approximately 50.0 μl of 1N NaOH/well to stop the reaction. Final absorbances were measured.

Provenance & Evidence