FreeStyle MAX システムは、哺乳類の一過性タンパク質産生用の完全なシステムです。このシステムは、Gibco FreeStyle 培地、FreeStyle MAX 試薬、および FreeStyle CHO-S 細胞または FreeStyle 293 細胞のいずれかを組み合わせたものです。FreeStyle MAX System を使用するために必要な装置は、CO2 インキュベーターと、振フラスコまたはスピナー培養を備えたプラットフォームシェイカーのみです。
FreeStyle MAX システムの特長: •浮遊培養における CHO および293細胞の一過性トランスフェクションのための簡単で迅速なプロトコル • 予想される翻訳後修飾を伴う最大ミリグラム量の機能性タンパク質 • FreeStyle™ による分泌タンパク質の単純化されたダウンストリーム精製 • 大規模生産のための簡単なプロトコル
FreeStyle MAX システムは、時間のかかる安定した細胞株の生成を、1週間で大規模な機能性タンパク質の生成に置き換えることができます。
What is the main difference between the FreeStyle Max 293 Expression System and the FreeStyle 293 Expression System?
The two systems have the same components except for the transfection reagent and the medium supplied for formation of transfection complexes. The FreeStyle Max 293 Expression System contains FreeStyle Max Reagent and OptiPRO SFM (serum-free medium that is devoid of animal-origin components), whereas the FreeStyle 293 Expression System contains 293fectin Transfection Reagent and Opti-MEM I Reduced Serum Medium (serum-free medium but not animal-origin free).
What is the recommended ratio of DNA and transfection reagent when transfecting Freestyle cells?
The optimal amount will vary, but we recommend starting with a 1:1 ratio of DNA and lipid. For example, 1.25 ug DNA and 1.25 ul of the reagent for one ml of suspension cells.
Which vectors do you recommend for use with FreeStyle system?
Generally, any mammalian expression vector should be compatible with the FreeStyle expression system. Some of the more popular choices include pcDNA3.3-TOPO TA vector (Cat. No. K8300-01), pcDNA3.4-TOPO TA vector (Cat. no. A14697), pcDNA3.2 DEST vector (Cat. No. 12489018), pCEP vector (Cat. No. V044-50), or pcDNA4/HisMax vector (Cat. No. K864-20).
How does FreeStyle MAX reagent compare to Lipofectamine 2000 and 293fectin reagents?
Lipofectamine 2000 is designed for adherent cells. FreeStyle and 293fectin reagents are designed for suspended Hek293 and CHO cells. For transfecting 293 suspension cells, FreeStyle MAX reagent yields transfection efficiency comparable to Lipofectamine 2000 and 293fectin reagents. However, Lipofectamine 2000 and 293fectin reagents do not work well for CHO suspension cells.