N2 Supplement Solution (100×)
| Reagent | Stock concentration | Volume for N2 (100×) | Concentration in N2 (100×) | Concentration in DMEM/F12 after 100× dilution |
|---|---|---|---|---|
| Apo-transferrin (Sigma-Aldrich T1147) | 100 mg/mL | 1 mL | 10 mg/mL | 100 µg/mL |
| BSA (Invitrogen 15260-037) | 75 mg/mL | 0.67 mL | 5 mg/mL | 50 µg/mL |
| Insulin (Sigma-Aldrich I1882) | 25 mg/mL | 1 mL | 2.5 mg/mL | 25 µg/mL |
| Progesterone (Sigma-Aldrich P8783) | 0.6 mg/mL | 0.033 mL | 0.002 mg/mL | 0.02 µg/mL |
| Putrescine (Sigma-Aldrich P5780) | 160 mg/mL | 0.1 mL | 1.6 mg/mL | 16 µg/mL |
| Sodium selenite (Sigma-Aldrich S5261) | 3 mm | 0.01 mL | 3 µm | 30 nm |
| DMEM/F12 | 7.187 mL | |||
| Store at 4°C and use within 1 mo. | ||||
