View CartMy Account / Order StatusHelp
:searchbox-only>
 

*TIP omit words like "with" or "and" from your search for better results


Join our mailing list!


(Your shopping cart is empty)
  Home
About Us
 Contact Us
 Careers  
You are here: Home > Protein Expression
We found 24 results matching your criteria. Narrow your search by selecting additional criteria below.
Choose a sub category:
Cinnabar Media Hi-Def Azure Media
Inducers Stable Isotopic Labeling Media
PROTEIN EXPRESSION

Teknova offers both Complex and Defined protein expression media for small-scale and large-scale protein expression in E. coli.
Cinnabar is a Complex Growth Media that reaches optical densities of OD600: 40-65
Hi-Def Azure is a Defined Growth Media, reaches optical densities of OD600: 9-14
Note: In comparison, LB OD600: 3-5 and TB OD600: 15-25

CINNABAR HIGH-YIELD PROTEIN EXPRESSION MEDIA
Teknova formulated Cinnabar Media, a complex growth medium, based on Studier's original formulation developed for protein expression in E. coli with a protein expression protocol developed by Studier known as auto-induction.
Cinnabar routinely reaches optical densities of OD600: 40-65
Compare to LB OD600: 3-5 and TB OD600: 15-25
Use of Cinnabar media can result in at least 3-8X more protein
Cinnabar media has been successfully used in E. colistrains BL21 and BL21(DE3).
We suggest supplementation of Cinnabar Media with 0.8% Glucose and 0.1mM IPTG.
Note: Cinnabar contains 0.7% Glycerol (so it is not necessary to supplement Cinnabar with Glycerol).

HI-DEF AZURE DEFINED GROWTH MEDIA
Teknova developed Hi-Def Azure Defined Growth Media based on Niedhardt’s original formulation for growth media for Enterobacteria
Hi-Def Azure can also be made in special formats for labeling and structural studies
Features of Hi-Def Azure Media :
1. All system components have a known chemical composition
2. Hi-Def Azure is Animal-Free
3. All components are tested in-house
4. Results in consistent performance
5. Eliminates lot-lot variability

PROTEIN EXPRESSION WITH AUTO-INDUCTION
A disadvantage of induction by IPTG is the need to monitor bacterial growth to achieve optimal induction conditions. This difficulty is especially apparent in experiments which seek to evaluate protein expression in a variety of clones. In this case, differences in growth rates will be observed and it will be unlikely to achieve optimal induction for all expression clones. The auto-induction protocol provides a method to circumvent these difficulties and simplify the expression protocol.

William Studier developed auto-induction as a method of protein expression in which cells are culture in media containing a mix of carbon sources, generally glucose, lactose and glycerol. Studier made a detailed study of auto-induction and found that in addition to simplifying the method for protein expression by simply allowing for addition of the inducer (typically IPTG) at the beginning of the protocol, Studier also found that by utilizing auto-induction, he was able to produce 5-20 times as much target protein per volume of culture as conventional IPTG induction.
In 2005, William Studier at Brookhaven introduced auto-induction as a simplified protocol for high-level protein expression in E. coli in T7-pET vectors.

Components in the media allow for the introduction of the IPTG inducer at the beginning of the experiment, thus eliminating the need for continual monitoring of cell growth. Due to catabolite repression, the presence of glucose in the media will serve to repress the inducer until at least the mid-logarithmic phase of growth, the optimal stage for high-level protein expression. When the glucose in the media is depleted, lactose (via IPTG) serves to induce expression of the T7 RNA polymerase upon conversion to allo-lactose by beta-galactosidase (the LacZ gene product). The media is well-buffered at all times and T7 RNA polymerase is induced in logarithmic to late-logarithmic phase of growth, which results in greater cell mass and higher yield of recombinant protein.

Studier points out that auto-induction should be performed only in strains encoding an intact lac operon. Studier's detailed study of auto-induction found that cell mass and total soluble target protein yield are increased several-fold using auto-induction when compared with conventional protein expression protocols.

ADVANTAGES of AUTO-INDUCTION
1. No need to monitor cell growth (IPTG is added at the beginning). final OD600 is at least 3X higher than with conventional IPTG induction protocol.
2. Greater cell mass will often result in greater soluble protein yield.
3. Can induce numerous expression clones simultaneously. (amenable for highthroughput).
4. Typically, lower amounts of IPTG are needed for induction (lowering the cost of protein expression).
5. Compatible with cultures grown in flasks, culture tubes and deep-well plates.
6. Minimal sample handling.

AUTO-INDUCTION PROTOCOL:
1. Transform plasmid into strain of E. coli optimized for protein expression. We use BL21 or BL21(DE3).
2. Inoculate 2 mL pre-culture for overnight growth:
A. Pick a single colony (from a freshly transformed plate) or
B. Use a thawed glycerol stock of cells to a final 1% (1:100) total culture volume (We add 20ul thawed glycerol stock into 2 mL Hi-Def Azure or Cinnabar, with appropriate antibiotic).
C. Grow pre-culture for 14-18 hours, shaking at 250-300rpm at 30-37oC in container with 5X volume for sufficient aeration. For example, for a 2mL pre-culture, we use a 13ml (17x100mm) polypropylene culture tube.

Note: It is common practice to prepare a pre-culture in advance and freeze aliquots as glycerol stocks at -70-80oC. These stocks should be thawed only once. Use of re-thawed glycerol stocks can result in >50% loss of viability with commensurate loss of maximum cell density.

3. Supplement Auto-induction Hi-Def Azure or Cinnabar media (at this time add appropriate antibiotic to media for selection).
For Hi-Def Azure media: we recommend adding 0.8% Glucose, 0.5% Glycerol and 0.1mM IPTG. For Cinnabar media: we recommend adding 0.8% Glucose and 0.1mM IPTG (Do not supplement Cinnabar with Glycerol; Cinnabar media already contains 0.7% Glycerol).

4. Inoculate media for auto-induction to 1% total culture volume (1:100).
Note: The size of the inoculum is a key variable to control for reproducibility. The size of the inoculum will affect the consistency of the results. We advise consistency in the size of the inoculum and consistency of the total inoculated volume to reduce variability between protein expression trials.

Induction will occur at an earlier time in the growth phase in 0.1% Glucose (than with 0.8% Glucose). We suggest supplementing with 0.8% Glucose to suppress the time of induction for toxic proteins or proteins that are known to reduce cell growth. For early auto-induction: Use 0.1% Glucose supplemented media. For late auto-induction: Use 0.8% Glucose supplemented media.

5. Grow culture at 16-30oC, shaking at 225-300 rpm for at least 22-28 hours for optimum protein expression.
We shake cultures at 300 rpm, (using Deutz microreactors) to increase aeration resulting in maximum cell density (OD600 = 50). For optimum protein expression, typically it is best to grow the cells well into stationary phase. Note: Cinnabar Media has a long lag-phase and will typically reach stationary phase at 26-28 hours.

6. Harvest Cells
Note: These are general guidelines. The best temperature for protein expression and the amount of time needed for optimum expression must be determined experimentally, for each protein. In general, for proper protein folding, for decreased production of inclusion bodies and for minimizing exposure to proteases, lower temperature of protein expression is suggested.

AUTO-INDUCTION REFERENCES:
1. Studier F. W. Protein Production by Autoinduction in High-Density Shaking Cultures. Protein Expression and Purification. 41(1):207-234.(2005). PMID: 15915565
2. Carnes A.E., Hodgson C.P., Williams J.P. Inducible Escherichia coli Fermentation for increased plasmid DNA production. Biotechnol. Appl. Biochem.; 45(3): 155-66.(2006). PMID: 16819941

TECHNICAL RESOURCES FOR PROTEIN EXPRESSION
Protein Expression Articles: References for Protein Expression
Poster by Teknova Scientists: Parameters Affecting Auto-Induction in Rich and Defined Media

Sort By:
1
G0565
100X Glucose, Glycerol, IPTG Auto Induction Supplement, Sterile. 1000mL Cat.No. G0565

Our Price: $50.94
G0560
100X Glucose, Glycerol, IPTG Auto-Induction Supplement, Sterile. 100mL Cat.No. G0560

Our Price: $33.96
G0575
100X Glucose, Glycerol, Lactose Auto-Induction Supplement, Sterile. 1000mL Cat.No. G0575

Our Price: $50.94
G0570
100X Glucose, Glycerol, Lactose Auto-Induction Supplement, Sterile. 100mL Cat.No. G0570

Our Price: $33.96
L0360
20% Lactose Solution, Sterile. 1000mL Cat.No. L0360

Our Price: $46.80
G9005
50% Glucose Solution. Sterile, 500ml. Cat.No. G9005

Our Price: $36.11
G1799
50% Glycerol Solution, 1000ml. Cat.No. G1799

Our Price: $66.62
G1800
50% Glycerol Solution, Animal Free, Sterile. Cat.No. G1800

Our Price: $52.01
G1796
50% Glycerol Solution. Sterile, 100ml Cat.No. G1796

Our Price: $34.37
G0580
50X Glucose, Glycerol, IPTG Auto Induction Supplement, Sterile. 100mL. Cat.No. G0580

Our Price: $33.96
2A7010
Autoinduction Media. 1 Liter. Sterile Cat. No. 2A7010

Our Price: $131.17
3H5081
Azure CF 50X Media for Fermentation Kit, Sterile. Cat.No. 3H5080

Product price is not filled in. This product cannot be purchased.

3H5080
Azure CF 50X Media for Fermentation Kit, Sterile. (3 part Kit - 1 Liter each) Cat.No. 3H5080

Our Price: $314.16
3S3800
Azure SSF. 1000mL, Sterile. Cat.No. 3S3800

Our Price: $46.08
B1842
Bradford Reagent (5X Concentrate). 1000mL, Sterile. Cat. No. B1842

Our Price: $141.75
B8105
Bradford Reagent (5X Concentrate). 500mL, Sterile. Cat. No. B8105

Our Price: $84.00
3C8488
Cinnabar High Yield Protein Expression Media, 1000mL, Sterile Cat.No. 3C8488

Our Price: $40.05
3H6200
Hi Def Azure Media Animal-Free, Sterile. 200mL in 1000mL Bottle Cat.No. 3H6200

Our Price: $44.57
3H5200
Hi Def Azure Media, Sterile. 200mL in 1000mL Bottle Cat.No. 3H5200

Our Price: $42.45
3H5002
Hi-Def Azure Media 1000mL, Sterile. A2010 20% Arabinose Solution, 50ml INCLUDED Cat.No. 3H5002

Our Price: $46.08
3H5000
Hi-Def Azure Media without Carbon source. Sterile. 1000mL Cat.No. 3H5000

Our Price: $46.08
3H5005
Hi-Def Azure Media, Sterile. 5 Liters Cat.No. 3H5005

Our Price: $285.00
3H5050
Hi-Def Azure Plus Media. 1 Liter Bottle, Sterile. Animal-Free. Cat.No. 3H5050

Our Price: $48.39
T5876
TY Base for Auto-Induction. 1 Liter Bottle, Sterile Cat.No. T5876

Our Price: $39.29
   
 
1
 

 About Us
 Contact Us
 Terms and Conditions
 Privacy Policy
 Send Us Feedback
 
Company Info| Product Index | Category Index | Help | Terms of Use
Copyright © www.teknova.com. All Rights Reserved. Order by Phone: 831-637-1100
Sell Online with Built with Volusion