Sucrose → HCID 19430206, compound

Participants

Reaction Steps

For over-expression of ATGL and CGI-58 in E. coli (XL-1) cDNA was cloned into the vector pASK-IBA5+ (IBA-BioTagnology). Cells were transformed and cultured over night at 30° C. Thereafter cells were transferred into a fresh medium and grown until OD600 reached 0.7-0.8. Expression was induced using 0.2 μg/ml anhydrotetracycline. After 3 hours incubation at 37° C. cells where harvested, resuspended in lysis buffer (0.25M Sucrose, 1 mM DTT, 1 mM EDTA) and disrupted by sonication. Lysates were centrifuged at 15,000 g for 20 min at 4° C. and the supernatant was collected.

Provenance & Evidence