Eukaryotic Expression and Purification of Recombinant Extracellular Matrix Proteins Carrying the Strep II Tag - Methods in Molecular Biology: Extracellular Matrix Protocols - page 50

Fig. 1. Vectors for the expression of secreted proteins with N- and C-terminal Step
tags in mammalian cells.

2. Growth media; serum-free media with 10% fetal calf serum ( not heat-inactivated

serum) and 175 µ g/mL active G418 (Gibco).

3. Selection media; serum-free media with 10% fetal calf serum, 175 µ g/mL active

G418 and 0.5 µ g/mL puromycin (Sigma, St. Louis, MO). ( See Note 1 .)

2.2. Materials for Purification of Protein
1. Protease inhibitors; X100 stock of NEM (N-ethylmaleimide, Fluka, Buchs, Swit-

zerland) and PMSF (phenylmethylsulfonyl fluoride, Fluka) are dissolved together
both at 100 m M in methanol and stored at -20 ° C.