Prokaryotic Expression
10X the protein yield from
your T7 expression system
MagicMedia™ E. coli Expression Medium
→ 3–10 times higher protein yields with much less hands-on time
→ Simple protocol eliminates tedious O.D. monitoring and induction steps
→ Compatible formulation works with 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 Invitrogen’s MagicMedia™ E. coli Expression Medium.
Superior protein yields
The MagicMedia™ formulation provides optimal conditions for
handling and scale-up processes. MagicMedia™ Medium is also
effectively regulating expression of the T7 promoter used in
ideal for efficiently performing simultaneous expression of many
many E. coli systems. This routinely results in high-density cul-
clones for high-throughput applications.
tures, producing recombinant protein yields that exceed those
IL-1
achieved using LB medium with IPTG inducer (Figure 1).
Ultimate convenience
MagicMedia™ E. coli Expression Medium is extremely easy to
use and does not require the time-consuming optical density
kDa
HPHLA2
JNK2
RacKin
Fatty
acid BP
HSU
18291
INT
ER beta
M 1 2 3 1 2 3 1 2 3 M 1 2 3 1 2 3 1 2 3 M 1 2 3 1 2 3
98
62
49
38
28
17
(O.D.) monitoring and induction steps used in conventional
LB + IPTG methods. 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
Figure 1—Higher protein yields achieved with the MagicMedia™ E. coli
Expression 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™ 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.
Prokaryotic Expression
Two all-inclusive formats:
powder and liquid
Traditional protocol—tedious multiple steps
MagicMedia™ E. coli Expression Medium
consists of proprietary components
Easy MagicMedia™ protocol
Step 1
Step 1
Pick colonies and
inoculate into LB
medium.
Pick colonies and
inoculate into LB
medium.
May skip step 1 for
culture volumes less
than 100 ml.
Step 2
A and B and is available in two
Inoculate LB medium for
expression.
convenient formats to meet specific
Step 2
Inoculate MagicMedia™
Medium for expression
overnight.
needs—premixed powder packaged
in a water-dissolvable bag (SoluPouch™
Steps 3–7
format) or a ready-to-use, presterilized
Check O.D. throughout
the day (3–7 times).
liquid. For the SoluPouch™ format, simply
place the entire pouch containing the
Step 8
component A powder in water, and
At proper O.D., add IPTG
inducer, grow 3–5 hours.
autoclave. The pouch and powder will
dissolve completely—the pouch will not
have any effect on the cells you culture.
Then add the liquid component B and
Step 9
Step 3
At the end of the day,
harvest and freeze pellet
for next day analysis.
At the beginning of the
day, harvest and analyze
expression results.
your antibiotic. There’s no weighing out
of 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
Figure 2—Effortless MagicMedia™ protocol.
Ordering information
Product
MagicMedia™ E. coli Expression Medium
Powder SoluPouch™ format
Ready-to-use liquid format
Quantity
1 L
5х1L
1L
Cat. no.
K6801
K6802
K6803
how easy it is to incorporate MagicMedia™
Medium into your current E. coli expres-
T7-regulated expression vectors and E. coli
sion procedures.
Champion™ pET300/NT-DEST and pET301/CT-DEST Gateway® Vector Kit
6 µg of each vector
K6300-01
Champion™ pET302/NT-His and pET303/CT-His Vector Kit
10 µg of each vector
K6302-03
BL21(DE3) Chemically Competent E. coli
20 × 50 µl
C6000-03
Related products (visit www.invitrogen.com/proteinexpression for more information)
Additional T7-regulated E. coli expression vectors
Protein purification reagents
www.invitrogen.com
©2006 Invitrogen Corporation. All rights reserved. These products may be covered by one or more Limited Use Label Licenses (see Invitrogen catalog or www.invitrogen.com). By use of these products you accept the
terms and conditions of all applicable Limited Use Label Licenses. For research use only. Not intended for any animal or human therapeutic or diagnostic use, unless otherwise stated. F-067239-r1 0906