pET28b-MPMVGag

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Large scale recombinant Pr78Gag-His6-tagged protein expression was performed by inoculating a single colony into 25 ml LB media containing kanamycin antibiotic (50 µg/ml), and cultured at 37 °C, overnight with shaking at 200 rounds per minute (rpm). The overnight culture was used to inoculate 500 ml LB media containing kanamycin (50 µg/ml) in a 2-liter baffled Erlenmeyer flask. The culture was grown at 28 °C until an OD600 nm of approximately 0.6 was reached. Protein expression was induced with the addition of 0.4 mM IPTG, and the cells were grown for a further 4 h at 28 °C.	Cells were harvested by centrifugation (6,300 g) at 4 °C for 15 minutes and cell pellets were stored frozen at −80 °C.

Protocol tips
Large scale recombinant Pr78Gag-His6-tagged protein expression was performed by inoculating a single colony into 25 ml LB media containing kanamycin antibiotic (50 µg/ml), and cultured at 37 °C, overnight with shaking at 200 rounds per minute (rpm). The overnight culture was used to inoculate 500 ml LB media containing kanamycin (50 µg/ml) in a 2-liter baffled Erlenmeyer flask. The culture was grown at 28 °C until an OD600 nm of approximately 0.6 was reached. Protein expression was induced with the addition of 0.4 mM IPTG, and the cells were grown for a further 4 h at 28 °C.
Downstream tips
Cells were harvested by centrifugation (6,300 g) at 4 °C for 15 minutes and cell pellets were stored frozen at −80 °C.

Publication protocol

Large scale recombinant Pr78Gag-His6-tagged protein expression was performed by inoculating a single colony into 25 ml LB media containing kanamycin antibiotic (50 µg/ml), and cultured at 37 °C, overnight with shaking at 200 rounds per minute (rpm). The overnight culture was used to inoculate 500 ml LB media containing kanamycin (50 µg/ml) in a 2-liter baffled Erlenmeyer flask. The culture was grown at 28 °C until an OD600 nm of approximately 0.6 was reached. Protein expression was induced with the addition of 0.4 mM IPTG, and the cells were grown for a further 4 h at 28 °C. Cells were harvested by centrifugation (6,300 g) at 4 °C for 15 minutes and cell pellets were stored frozen at −80 °C.

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

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Protein Expression Prokaryotic cells - E. coli (MPMV) Pr78Gag using pET28b-MPMVGag from Tahir Aziz Rizvi, Department of Microbiology & Immunology, Colle.

Manufacturer protocol

Download the product protocol from Tahir Aziz Rizvi, Department of Microbiology & Immunology, Colle for pET28b-MPMVGag below.

Videos

Check out videos that might be relevant for performing Protein Expression Prokaryotic cells - E. coli (MPMV) Pr78Gag using pET28b-MPMVGag from Tahir Aziz Rizvi, Department of Microbiology & Immunology, Colle. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.