pMJS205

Protein Expression Prokaryotic cells - E. coli S. cerevisiae SOX Erv1p

Experiment
Protein Expression Prokaryotic cells - E. coli S. cerevisiae SOX Erv1p
Product
pMJS205 from Robyn Roth, Biosciences, Council for Scientific and Industrial R
Manufacturer
Robyn Roth, Biosciences, Council for Scientific and Industrial R

Protocol tips

Protocol tips
For protein expression, precultures were prepared by inoculating the recombinant strains in 5 ml LB (10 g l−1 tryptone, 5 g l−1 yeast extract, 10 g l−1 NaCl) medium containing kanamycin (50 mg l−1) and/or chloramphenicol (34 mg l−1), depending on plasmids present, and cultivated overnight at 37°C. A 1 in 100 dilution was made into 50 ml fresh LB media and cultivation continued at 30°C until OD600 = 0·6, followed by induction with 0·5 mol l−1 isopropyl‐β‐d‐thiogalactopyranoside (IPTG).

Publication protocol

For protein expression, precultures were prepared by inoculating the recombinant strains in 5 ml LB (10 g l−1 tryptone, 5 g l−1 yeast extract, 10 g l−1 NaCl) medium containing kanamycin (50 mg l−1) and/or chloramphenicol (34 mg l−1), depending on plasmids present, and cultivated overnight at 37°C. A 1 in 100 dilution was made into 50 ml fresh LB media and cultivation continued at 30°C until OD600 = 0·6, followed by induction with 0·5 mol l−1 isopropyl‐β‐d‐thiogalactopyranoside (IPTG). The cell pellets were harvested by centrifugation 20 h after induction, then resuspended in an equivalent volume Binding Buffer (20 mmol l−1 Tris pH 7·9, 500 mmol l−1 NaCl, 5 mmol l−1 imidazole) and sonicated for 5 min on continuous pulse using a sonicator (SonoPuls, Bandelin Electronic, Berlin, Germany) at 67% power. Centrifugation for 10 min at 9168 g yielded supernatant (soluble fraction) and pellet (insoluble fraction). The pellet was again resuspended in an equivalent volume Binding Buffer. Soluble and insoluble fractions were visualized on 12% SDS‐PAGE gels.

Full paper   Login or join for free to view the full paper.

Reviews

pMJS205 from Robyn Roth, Biosciences, Council for Scientific and Industrial R has not yet been reviewed for this experiment

We'd love it if you would be the first to write a review!

Discussion

Start your discussion

Share your thoughts or question with experts in your field

Start a discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Protein Expression Prokaryotic cells - E. coli S. cerevisiae SOX Erv1p using pMJS205 from Robyn Roth, Biosciences, Council for Scientific and Industrial R.

View full paper   Login or join for free to view the full paper.

Manufacturer protocol

Download the product protocol from Robyn Roth, Biosciences, Council for Scientific and Industrial R for pMJS205 below.

We haven't found the manufacturer protocol for this product yet.

Videos

Check out videos that might be relevant for performing Protein Expression Prokaryotic cells - E. coli S. cerevisiae SOX Erv1p using pMJS205 from Robyn Roth, Biosciences, Council for Scientific and Industrial R. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.

We haven't found any additional videos for this experiment / product combination yet.

Outsource your experiment

Fill out your contact details and receive price quotes in your Inbox

  Outsource experiment
Become shareholder Discussions About us Contact Privacy Terms