Get improved protein yield from your T7 expression clones
- 3–10 times higher protein yields with much less hands-on time
- Simple protocol eliminates tedious OD monitoring and induction steps
- Compatible formulation works with all T7-regulated E. coli vectors and BL21 and derivative strains
Due to their ease of use, relatively high yields, and simple scalability, T7-regulated E. coli expression systems and BL21 hosts are often chosen for generating recombinant protein. Now you can further increase yields and make this already easy expression system even easier-simply by switching to MagicMedia™ E. coli Expression Medium.
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
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.
For Research Use Only. Not for use in diagnostic procedures.