PMC:101390 / 27538-28399 JSONTXT

{"project":"2_test","denotations":[{"id":"11914155-9210347-8561989","span":{"begin":85,"end":86},"obj":"9210347"},{"id":"11914155-942051-8561990","span":{"begin":622,"end":624},"obj":"942051"}],"text":"The extraction of cell-linked COX by using Triton X-100 was as described previously [9]. For the cell linked COX extraction and purification by the Triton X-114 method, the extract obtained in cold after removal of cells by centrifugation was submitted to temperature-induced phase separation. The coalescence of the detergent was facilitated by warming up to 37°C for 15 min that was followed by the sharp separation of the two resulting phases by spinning at 4000 g for 15 min at 25°C. Both phases, the lower detergent-rich and the upper detergent-depleted were assayed for both enzyme activity (see below) and protein [32]. Using Triton X-114 also purified the extracellular COX. The cold culture broth was supplemented with Triton X-114 to the desired final concentration and detergent was completely dissolved at 4°C. Phase separation was induced as above."}