Answer
Post centrifugation, the supernatant contains the soluble part of protein and pellet contains the insoluble parts of the proteins. Thus, the target protein comes in the supernatant post salti-precipitation.
(a) Upon additions of this supernatant in the cell culture, the protien tends to maintain its physical 3-D structure. To accomplish it, the protiens tends to accumulate all the salts present in the media to reach a high-salt content at its periphery so the at its tertiary structure is maintained. Since cell-culture requires a critical amount of ionic balance in the culture media, lack of optimum amount of salts causes cell death.
(b) To circumvent this problem, either a diluted form of the supernatant could be introduced in the culture media or the cell could be made adaptive to this absolute concentration of protein by growing them in serial dilution (incrasing order) of protien concentrations for 5-6 passages. This will make cells more adaptive towards high salt concentrations.
Work Step by Step
Post centrifugation, the supernatant contains the soluble part of protein and pellet contains the insoluble parts of the proteins. Thus, the target protein comes in the supernatant post salti-precipitation.
(a) Upon additions of this supernatant in the cell culture, the protien tends to maintain its physical 3-D structure. To accomplish it, the protiens tends to accumulate all the salts present in the media to reach a high-salt content at its periphery so the at its tertiary structure is maintained. Since cell-culture requires a critical amount of ionic balance in the culture media, lack of optimum amount of salts causes cell death.
(b) To circumvent this problem, either a diluted form of the supernatant could be introduced in the culture media or the cell could be made adaptive to this absolute concentration of protein by growing them in serial dilution (incrasing order) of protien concentrations for 5-6 passages. This will make cells more adaptive towards high salt concentrations.
