Why Does Ammonium Sulfate Saturation in Supernatant Vary Among Different Groups?

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SUMMARY

The discussion centers on the variability in ammonium sulfate saturation in supernatants among different student groups conducting protein purification. Despite starting with identical 10,000 x g supernatants and performing 40-65% ammonium sulfate cuts, results differ significantly. Key factors contributing to this variance include potential discrepancies in the measurement of ammonium sulfate and the rate of its addition to the supernatant, which can affect the saturation process and the resulting yield of purified proteins.

PREREQUISITES
  • Understanding of protein purification techniques
  • Knowledge of ammonium sulfate precipitation
  • Familiarity with enzyme purification processes
  • Basic laboratory skills in measuring and mixing solutions
NEXT STEPS
  • Research the principles of ammonium sulfate precipitation in protein purification
  • Learn about the effects of solution mixing rates on saturation levels
  • Explore methods for accurately measuring solid reagents in laboratory settings
  • Investigate the role of protein aggregation in enzyme purification
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This discussion is beneficial for students and researchers involved in biochemistry, particularly those focusing on protein purification methods and enzyme studies.

Violagirl
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Homework Statement



If all students start with the same amount of 10,000 x g supernatant, and after different groups of students have done their 40-65% ammonium sulfate cut, their results tend to vary greatly. What are likely reasons for this variance?


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The Attempt at a Solution



I was thinking that if everyone started out with the same amount of supernatant and if the ammonium sulfate needed to be measured out based on the volume of supernatant present, maybe it's possible that different amounts of ammonium sulfate could have been measured out in the beginning before it was added to the supernatant. Otherwise maybe that it's possible that the rate of speed at which the amount of ammonium sulfate added to the supernatant may have been different which could impact the rate at which the supernatant becomes saturated? Any other input on this would be great.
 
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Violagirl said:

Homework Statement



If all students start with the same amount of 10,000 x g supernatant, and after different groups of students have done their 40-65% ammonium sulfate cut, their results tend to vary greatly. What are likely reasons for this variance?

No idea what you are talking about. And I am not going to try to guess, please elaborate.
 
The question relates to a crude sample of enzyme that is in the process of becoming purified. The ammonium sulfate is being added to pull extra molecules off the proteins so that they will aggregate together and come out of the solution. In this case, it looks as if they want to give consideration to how saturation plays a part in how much of the supernatant a person will get and how/why variation occurs between the amount that each person will get based on how much ammonium sulfate is added into the solution.

I was not for sure on why one person may end up with a different amount of supernatant from another. Could it be that the rate at which the ammonium sulfate, after it is weighted out, is added to the supernatant could impact the rate at which saturation of the supernatant takes place?
 

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