反应详情

Anti-skatole antibodies were prepared essentially in the same manner as described by Weiler, Planta (1981) 153:319-324 for preparing antibodies to detect a plant hormone, indole acetic acid. A skatole analogue-, indole-3-acetic acid (IAA), bovine serum albumin (BSA) conjugate was prepared by a mixed anhydride method. IAA (52.3 mg, 300 μmol) in 2 ml dimethylformamide (DMF) was treated with 75 μl tri-n-butyl amine and the solution was cooled to about -10° to -15° C. Forty μl of isobutylchlorocarbonate was added to that mixture, and the solution was stirred for about 10 min. That mixture was continually stirred while a total of 420 mg of BSA, dissolved in a mixture of 23 ml 50% DMF solution and 420 μl of 1N NaOH was slowly added. Following addition of the BSA, the solution was stirred at 0° C. for an hour. 100 μl of 1N NaOH was then added. The mixture was stirred for an additional four hours at 0° C. That mixture was dialyzed twice against 2 L of 10% DMF and then against water (4×1 L).

1. customas described by Weiler, Planta (1981) 153:319-324 for preparing antibodies
2. temperaturethe solution was cooled to about -10° to -15° C
3. stirringThat mixture was continually stirred while a total of 420 mg of BSA
4. additionwas slowly added
5. additionaddition of the BSA
6. stirringthe solution was stirred at 0° C. for an hour
7. stirringThe mixture was stirred for an additional four hours at 0° C
8. customThat mixture was dialyzed twice against 2 L of 10% DMF