Product Data Sheet ExpressMax Fermentation Media.

Item Description

ExpressMax aging media help in the development of recombinant proteins. The series is included eight pre-planned media, intended to cover ascope of conceivable articulation conditions and amplify the development of recombinant proteins.

The ExpressMax basal salts are inorganic supplements enhanced to produce the most elevated conceivable development pace of E. coli , bringing about maximal creation of recombinant proteins.

The screening pack is expected to support plan improvement by providing the basal salts/minor components blend alongside the carbon source (glucose) and nitrogen sources (yeast extricate, soy protein hydrolysate and Atholate) which can be consolidated in diff erent varieties to distinguish the arrangement that gives the most noteworthy creation of the objective protein.

Directions for Use

1. Materials – Kit contains an adequate number of clean fluid reagents to lead one 8-jar
screening test
1.1. LB Broth
1.2. 5x Fermentation Salts and Trace Metals Mix (191.36 g/L)
1.3. 5x Yeast Extract (25 g/L)
1.4. 5x Atholate™ (25 g/L)
1.5. 5x Soy Hydrolysate (25 g/L)
1.6. 20x Glucose Nutrient Mix (200 g/L)
1.7. 250 mL astound lined jars (8)
1.8. Wash Buffer: 40 mM sodium phosphate pH 7.5, 150 mM NaCl
1.9. 5x SDS-PAGE Loading Dye: 124 mM Tris-Cl, 1.28 M 2-Mercaptoethanol,
0.1 M glycerol, 0.14 M SDS (sodium dodecyl sulfate),
2.9 mM bromophenol blue
1.10. Tris-Glycine SDS-polyacrylamide gel of proper piece.

2. Strategies
2.1. Immunize a solitary state of the recombinant strain into 10 mL of LB Broth
in a confuse lined shake carafe. Hatch at 37°C short-term.
2.2. Get ready eight 250 mL confuse lined carafes containing 25 mL of each of
the equations according to the table above.
2.3. Vaccinate 25 mL of every one of the eight media with 1 mL of the short-term
culture. Brood the way of life at 37°C until the OD600 comes to 0.6.
2.4. Eliminate a 1 mL (“pre-acceptance”) test, reap the cells in a microfuge
tube, and suspend the cells in water to a thickness of 10 OD/ml. Store
at – 80°C.
2.5. Add inducer and keep hatching for 3 hours.
2.6. Eliminate a 1 mL (“post-acceptance”) test and cycle as in sync 2.4.
2.7. Collect the rest of the way of life, wash the pellets with 10 mL of wash
support, reharvest the cells, disposing of the supernatant/wash. Decide the
mass of the cell pellet and store the cell pellets at – 80°C.
2.8. Dissect for articulation of the objective protein as follows:
2.8.1. To decide protein creation per mL of culture:
2.8.1.1. Defrost the resuspended cells from pre-and post-acceptance
at 37°C.
2.8.1.2. Blend 5 µL of every cell suspension with 35 µL water and
10 µL 5x SDS-PAGE stacking color. Heat at 100°C for 5 minutes
furthermore, load 10 µL per path of polyacrylamide gel.
2.8.2. Stain the gel with Coomassie Blue, colloidal Coomassie Blue or Silver
stain as the need should arise.

3. Understanding
3.1. Subsequent to staining the gel, notice every path and analyze the “pre-acceptance”
test with the “post-enlistment” test from every medium. Raised
articulation is shown by the presence of a novel polypeptide band
relating to the sub-atomic mass of the objective protein in the
“post-enlistment” test.
3.1.1. Look at the degree of target protein from cells filled in each of the
eight definitions of ExpressMax media. Select the medium
which creates the most elevated level of target protein per mL of
culture.
3.2. When the fitting nitrogen sources have been distinguished, further
advancement ought to be performed to decide the appropriate focus
of the nitrogen source(s) for the protein of interest.

Material Safety Data

FOR RESEARCH USE ONLY. NOT INTENDED OR APPROVED FOR HUMAN, DIAGNOSTICS OR VETERINARY
USE.

Try not to ingest, swallow or breathe in. Try not to get in eyes, on skin, or on apparel. Wash completely in the wake of taking care of. For complete wellbeing data see full Material Safety Data Sheet.

Individual Reagent Information

All pack parts (i.e., Atholate, Yeast Extract,Basal Salts, Glucose Nutrient Mix, and Soy Hydrolysate), as well as any of the 8 fi nal definitions are accessible in bigger amounts from AthenaES.

Bile salts
GX6607-100G	Glentham Life Sciences	100 g	151.2 EUR
Bile salts
GX6607-10G	Glentham Life Sciences	10 g	58.8 EUR
Bile salts
GX6607-250G	Glentham Life Sciences	250 g	255.6 EUR
Bile salts
GX6607-25G	Glentham Life Sciences	25 g	79.2 EUR
Bile salts
GX6607-50G	Glentham Life Sciences	50 g	103.2 EUR
Dehydration Salts
M-CO-121	MiTeGen	12 solutions	142 EUR
Kinesis Tub PEEK Red .0145 x 100um
CHR4765	Scientific Laboratory Supplies	1INCH	3.42 EUR
Earle's Balanced Salts
CA006-050	GenDepot	500ml	84 EUR
Earle's Balanced Salts
CA006-300	GenDepot	6x500ml	174 EUR
BILE SALTS #3
B02-126-10kg	Alphabiosciences	10 kg	4298.4 EUR
BILE SALTS #3
B02-126-2Kg	Alphabiosciences	2 Kg	981.6 EUR
BILE SALTS #3
B02-126-500g	Alphabiosciences	500 g	310.8 EUR
JBScreen Plus Salts
M-CS-503	MiTeGen	24 solutions	226 EUR
Chiller for Electrophoresis systems
CSLCHILLER	Scientific Laboratory Supplies	EACH	6453.54 EUR
Kinesis Tub PEEK Yel .0145 x 150um ONE PIECE
CHR4767	Scientific Laboratory Supplies	1INCH	2.28 EUR
MEM With Hank's Salts
CM050-050	GenDepot	500ml	92.4 EUR
MEM With Hank's Salts
CM050-300	GenDepot	6x500ml	199.2 EUR

Material Safety Data Sheet.

FOR RESEARCH USE ONLY. NOT INTENDED OR APPROVED FOR HUMAN, DIAGNOSTICS OR VETERINARY USE. Try not to ingest, swallow or breathe in. Try not to get in eyes, on skin, or on apparel. Wash completely in the wake of dealing with. For complete wellbeing data
see full Material Safety Data Sheet.