I have the following question on the amount of ammonium sulfate and
magnesium sulfate I would use for the following procedure I found in a
physiological chemistry book. I am working with one liter volumes or 10
gram solid weights. I am not familiar working with saturated solutions
of these two compounds. Thanks Kevin.
1. Immerse tissue in cold 0.25N sulfuric acid.
2. Drain off acid, mince tissue and suspend for 24 hours in 2 volumes of
0.25N sulfuric acid at 5 degrees C.
3. Strain through gauze.
4. Add solid ammonium sulfate to 0.4 saturation. How many grams of
ammonium sulfate do I add to 1 liter of solution?
5. Filter.
6. Saturate to 0.7 saturation. How many grams of ammonium sulfate do
I add to 1 liter of solution?
7. Dissolve the precipitate from 0.7 saturation in 0.25 saturated
ammonium sulfate. How many grams of ammonium sulfate do I add to
make a 0.25 saturated ammonium sulfate if I have 10 grams of precipt?
8. Adjust pH to 5.0.
9. Let stand 2 days at 25 degrees C.
10. Filter.
11. Recrystallize the crystals on the filter pad in 0.25 saturated
ammonium sulfate at pH 5.0 about 8 times. How many grams of
ammonium sulfate do I add to make a 0.25 saturated ammonium
sulfate if I have 10 grams of crystals?
12. Dissolve in 0.02N sulfuric acid.
13. Adjust pH to 4.0.
14. Salt out in 0.7 saturated ammonium sulfate. How many grams of
ammonium sulfate do I add to make a 0.7 saturated ammonium
sulfate to 1 liter of solution?
15. Filter.
16. Dissolve in 0.01N sulfuric acid.
17. Let stand for 24 hours at 25 degrees C.
18. Filter and save crystals.
19. Adjust filtrate from 18 to pH 4.0.
20. Precipitate in 0.7 saturated ammonium sulfate and refractionate
between 0.4 and 0.7 saturated ammonium sufate. What is meant
by this and what amounts in grams of ammonium sulfate do I add
say to 1 liter of solution?
21. Filter.
22. Wash precipitate from 0.7 saturation with saturated magnesium
sulfate annhydrous. How many grams of magnesium sulfate do I add
to make a saturated magnesium sulfate solution for 1 liter of
solution?
23. Dissolve in 0.4M borate at pH 9.0.
24. Cool to 5 degrees C.
25. Bring to 0.5 saturation with magnesium sulfate annhydrous. How
many grams of magnesium sulfate do I add to 1 liter?
26. Let stand 3 days at 5 degrees C.
27. Filter off the crystals.
28. Wash crystals in 0.5 saturated magnesium sulfate annhydrous. How
many grams of magnesium sulfate do I add to say 10 grams of
crystals?
29. Dissolve in 0.02N sulfuric acid.
30. Bring to 0.5 saturation with magnesium sulfate? How many grams do
I need of magnesium sulfate for 1 liter solution?
31. Add 0.4M borate to give a final pH of 9.0.
32. Let stand 1 day at 5 degrees C.
33. Filter and retain crystals.
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