Amide Coupling Array
Carbodiimide Coupling Reagent Kit (HCK1005-01-002)
2 sets of 6 reagents; HATU, HBTU, HCTU, COMU®, HDMC, HDMA, and ethyl(
The typical protocol is performed at 0.1 mol/L concentration reaction condition using a solution of combined acid and amine.
Protocol at 100 µl volume reaction condition
requiredvolume of a solution of acid substrate 0.2 mol/L in the desired solvent. For example, 350 µl solution for 6 reaction conditions (50 µl extra to compensate potential evaporation).
- Using a clean and dry syringe, add 50 µl of the acid solution to each reaction vial containing a coupling reagent. Stir the reaction vials in the reaction block at
desiredtemperature (between 0°C and 25°C) for 30 min.
- Prepare 350 µl of a solution of amine substrate 0.2 mol/L and base (Et3N or DIPEA) 0.4 mol/L in the desired solvent. Add 50 µl of this solution to each reaction vial.
- Optional: Add 10 µl of the solution of ethyl (
hydroxyimino) cyanoacetate to each reaction vial. (prepared by adding 70 µl of DMF in the reagent vial)
- Stir the reaction vials in the reaction block at
desiredtemperature for 3 to 12 hours. Do not heat vials at temperaturehigher than 120°C or higher than 5°C above the boiling point of the used solvent.
- Remove the vial caps using a
analyticalsample for each reaction condition with 5 µl sample diluted into 200 µl in either DMSO or water/acetonitrile 50/50. Analyze resulting analytical samples by LC/MS.
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