Add to DMEM/F12 medium the following additives
|
ingredients |
Stock reagents |
Vol.
in 10 ml |
Final conc. |
Expiration 4°
C -20° C |
|
Insulin |
100 mg/ml |
25 ml |
250 ng/ml |
1 mo |
5 mo |
|
Transferrin |
20 mg/ml |
5 ml |
10 mg/ml |
1 mo |
3 mo |
|
Sodium
Selenite |
2.6 mg/ml |
10 ml |
2.6 ng/ml |
1 wk |
1 mo |
|
Estradiol |
10-7 M |
10 ml |
1010 M |
3 mo |
6 mo |
|
Hydrocortisone |
1.4x10-3 M |
10 ml |
1.4x10-6 M |
1 mo |
6 mo |
|
Prolactin |
1 mg/ml (30u/ml) |
50 ml |
5 mg/ml |
1 mo |
6 mo |
|
EGF (S1
only) |
20 mg/ml |
5 ml |
10 ng/ml |
2 wk |
3 mo |
The additives we supplement DMEM/F12 media (GibcoBRL #12400-024) with to produce H14 medium are purchased from the following vendors. Please note that EGF is only for H14 medium for S1 cells; T4-2 cells do not require EGF. The paper that describes how to made the media additives is entitled “Cell Differentiation by Extracellular Matrix Components” by R. J. Blaschke, A. R. Howlett, P. Deprez, O. W. Petersen, and M. J. Bissell, found in Methods in Enzymology 245:535-569. If you cannot obtain a copy, we can fax one to you.
Ingredients Vendor Catalog number
Insulin Sigma I-6634
Transferrin (Human) Sigma T-2252
Sodium Selenite Collab.
Research 40201
Estradiol Sigma E-2758
Hydrocortisone Sigma H-0888
Prolactin Sigma L-6520
(or can obtain from National Institute of Diabetes
and Digestive and Kidney Disease (NIDDK))
EGF Collab. Research 40001