Protocol for CEPT Cocktail Kit
This is intended as a guide only.
In Brief Download the PDF of this protocol
CEPT Cocktail is a combination of small molecules, Chroman 1 (Cat. No. 7163), Emricasan (Cat. No. 7310), Polyamine Supplement x1000 (lyophilized) (Cat. No. 7739), and trans-ISRIB (Cat. No. 5284) available from Tocris, which improves the viability of pluripotent stem cells (PSCs). This combination has been shown to be effective in cryopreservation, routine long-term passaging, as well as embryoid body and organoid formation.
CEPT Cocktail Kit
| Product |
Catalog Number |
Unit Size |
Stock Solution Concentration |
Volume for Reconstitution |
|---|---|---|---|---|
| Chroman 1 | 7163 | 1 vial | 10,000x | 500 μL DMSO for a 0.5 mM solution |
| Emricasan | 7310 | 1 vial | 10,000x | 500 μL DMSO for a 50 mM solution |
| trans-ISRIB | 5284 | 1 vial | 10,000x | 500 μL DMSO for a 7 mM solution |
| Polyamine Supplement x1000 (lyophilized) |
7739 | 1 vial | 1000x | 5 mL ddH2O |
| DMSO, sterile filtered | 3176 | 5 mL |
Reconstitution of stock solutions
1. Prepare stock solutions (10,000x) for Chroman 1 (0.5 mM), Emricasan (50 mM) and trans-ISRIB (7 mM) by adding 500 μL of DMSO, sterile filtered, to each vial. Mix thoroughly or gently warm the solutions in a 45-60ºC water bath to ensure complete reconstitution. Add 100 μL of resuspended 10,000x stock solution per 1 L of media.
2. Prepare a 1000x Polyamine Supplement stock solution by reconstituting vial contents in 5 mL of ddH2O and filter sterilize before use. Add 1 mL of sterile resuspended polyamines supplement (1000x stock) per 1 L of media.
3. Filter the prepared medium before use.
Storage and Handling
Store the stock solutions, once prepared, at -80°C and use them within 6 months, or at -20°C within 1 month.