Dual Media Set™ (EB + OB)
About Dual Media Set™ (EB + OB)
Although recombinant protein expression in E. Coli has become routine, high level protein expression or overexpression is not always attainable for every protein. Our research has shown that high level protein expression can be achieved consistently when two processes, cell expansion and protein expression, are kept separate. The Dual Media Set™, different from commonly used protein expression procedures using Luria-Bertani (LB) medium or other specially prepared medium, contains two specially formulated media: Expansion Broth (EB) and Overexpression Broth (OB). For expansion, E.coli cells are grown in EB which keeps the production of recombinant protein repressed. To initiate high level protein expression, OB is simply added to the culture. By using the Dual Media Set™, protein overexpression can be reliably controlled for many recombinant proteins (see Figure 1 below). In some circumstances, when the expressed protein is either toxic or insoluble, overexpression may be counter-productive. In such cases, protein production can be kept at a minimum by adding the inducer IPTG (for lac-based promoters) to cells growing in EB (see Figure 2 below).
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FIGURE 1: SDS-PAGE of cell proteins after growth using the Dual Media Set™. M "“ protein markers;1-5, West Nile virus protein E (54 kDa): 1, repressed expression in EB, 2-5, over-expression in OB for 5, 10, 18, and 24 hours, respectively, after inoculation with uninduced EB culture; 6-10, C-terminal domain of West Nile virus protein E (32 kDa): 6, repressed expression in EB, 7-10, over-expression in OB for 5, 10, 18, and 24 hours, respectively, after inoculation with uninduced EB culture; 11-15, Methyl-binding domain of MeCP2 (18 kDa): 11, repressed expression in EB, 12-15, over-expression in OB for 5, 10, 18, and 24 hours, respectively, after inoculation with uninduced EB culture.
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FIGURE 2: Controlled overexpression of ß-galactosidase. Cells were grown in EB, where only background levels of the T7-lac promoter-controlled product are produced (1). Moderate amounts of the enzyme were produced by incubating overnight in EB with IPTG (2), the highest amounts of protein are produced in OB (3).
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Simple, reliable method for high level recombinant protein expression in E. coli.
Eliminates the need to monitor cell density and the time of inducer addition.
Synchronizes cultures that express different recombinant proteins.
| Product |
Size |
Catalog # |
How to Buy |
| Dual Media Set™ (EB + OB) |
100ml EB™-500ml OB™ |
M3011 |
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