使用你的T7表达克隆获取更高的蛋白得率

  • 花费少得多的手动操作时间,同时获得3-10倍高的蛋白得率
  • 实验方案简单,消除了冗长的OD监控及诱导步骤
  • 兼容配方可用于所有的T7调控的大肠杆菌载体及BL21和衍生菌株

T7调控的大肠杆菌表达系统及BL21宿主使用简单、得率相对较高、培养规模简单可调,常被选用来制备重组蛋白质。现在你可以进一步提高得率,并使这一已经很简单的表达系统变得更加简单——只需要换用MagicMedia™ 大肠杆菌表达培养基即可。

Superior protein yields

The MagicMedia™ formulation provides optimal conditions for effectively regulating expression of the T7 promoter used in manyE. coli systems. This routinely results in high-density cultures that produce high recombinant protein yields that exceed those achieved using LB medium supplemented with IPTG inducer (Figure 1).

Figure 1.  Increase protein yields with the MagicMedia™ E. coli medium. 

Eight different human ORFs were cloned into the Champion™ pET300/NT-DEST vector using Gateway® cloning. Positive clones were transformed into BL21(DE3) E. coli. The clones were expressed in LB + IPTG (lane 1), ready-to-use liquid MagicMedia™ medium (lane 2), or MagicMedia™ medium prepared from powder (SoluPouch™ format) (lane 3). Two hundred microliters of each culture was lysed and analyzed on a Coomassie®-stained NuPAGE® Novex® 4–12% Bis-Tris Gel. M: SeeBlue® Protein Standard

Ultimate convenience

MagicMedia™ E. coli Expression Medium is extremely easy-to-use and does not require the time-consuming optical density (OD) monitoring and induction steps as conventional LB + IPTG methods do. Simply inoculate prepared MagicMedia™ medium with the appropriate seed culture, grow the culture overnight, and harvest (Figure 2). In addition, the E. coli culture volume required when using the MagicMedia™ formulation is significantly reduced compared to LB volumes, simplifying handling and scale-up processes. MagicMedia™ medium is also ideal for efficiently performing simultaneous expression of many clones for high-throughput applications.

Figure 2.  Effortless MagicMedia™ protocol. 

Two all-inclusive formats: powder and liquid

MagicMedia™ E. coli Expression Medium consists of proprietary components A and B and is available in two convenient formats to meet specific needs-premixed powder packaged in a convenient, water-dissolvable bag (SoluPouch™ format) or a ready-to-use, presterilized liquid. For the SoluPouch™ format, simply place the entire pouch containing the component A powder in water and autoclave. The SoluPouch™ and powder will dissolve completely—the pouch will not have any affect on the cells you culture. Then add the liquid component B and your antibiotic. There's no weighing out materials, no mess, and no additional reagents needed (except your selection agent). For the ready-to-go liquid format, simply combine components A and B and add your antibiotic-no autoclaving is necessary. With either format, you'll see how easy it is to incorporate MagicMedia™ Medium into your current E. coli expression procedures.