Topic: Isolation of Crude Ovalbumin from Egg White by Ammonium Sulfate Precipitation (Salting Out) crude ovalbumin was isolated from egg whites of two medium-sized eggs. Egg whites contain about 88.5% water and about 10% proteins, from which ovalbumin is the major component. The eggs were cracked open, and the egg whites were separated carefully from the yolk. Stirring of the egg whites was done to break up the membrane. This was followed by filtration through cheesecloth, where a clear filtrate was obtained. About 40.0 mL of the filtered undiluted egg white was measured, and 0.10 mL of 1 M acetic acid was slowly added for every 1.0 mL of egg white with constant stirring. A turbid mixture with few amounts of white precipitate may be expected to form. These may be removed by filtering the mixture using a cheesecloth. After filtration, 30.0 mL of the filtrate was obtained, and the solution was brought from 0% to 40% saturation by adding the required amount of powdered ammonium sulfate, (NH4)2SO4. In calculating the required amount of powdered (NH4)2SO4, an ammonium sulfate saturation table is needed The computed amount of powdered ammonium sulfate was added to the egg white sample portion by portion with constant stirring while submerged in an ice bath. The solution is expected to become more turbid, and a white precipitate is expected to form The resulting mixture was then filtered using a cheesecloth. The residue was discarded, and 30.0 mL of the filtrate was brought from 40% to 60% saturation by adding the required amount of powdered ammonium sulfate in the same manner as the previous addition. QUESTION: Give the purpose of bringing the solution to 0-40% ammonium sulfate saturation and then 40-60% saturation.
Topic: Isolation of Crude Ovalbumin from Egg White by Ammonium Sulfate Precipitation (Salting Out)
crude ovalbumin was isolated from egg whites of two medium-sized eggs. Egg whites contain about 88.5% water and about 10% proteins, from which ovalbumin is the major component. The eggs were cracked open, and the egg whites were separated carefully from the yolk. Stirring of the egg whites was done to break up the membrane. This was followed by filtration through cheesecloth, where a clear filtrate was obtained. About 40.0 mL of the filtered undiluted egg white was measured, and 0.10 mL of 1 M acetic acid was slowly added for every 1.0 mL of egg white with constant stirring. A turbid mixture with few amounts of white precipitate may be expected to form. These may be removed by filtering the mixture using a cheesecloth.
After filtration, 30.0 mL of the filtrate was obtained, and the solution was brought from 0% to 40% saturation by adding the required amount of powdered ammonium sulfate, (NH4)2SO4. In calculating the required amount of powdered (NH4)2SO4, an ammonium sulfate saturation table is needed
The computed amount of powdered ammonium sulfate was added to the egg white sample portion by portion with constant stirring while submerged in an ice bath. The solution is expected to become more turbid, and a white precipitate is expected to form
The resulting mixture was then filtered using a cheesecloth. The residue was discarded, and 30.0 mL of the filtrate was brought from 40% to 60% saturation by adding the required amount of powdered ammonium sulfate in the same manner as the previous addition.
QUESTION:
Give the purpose of bringing the solution to 0-40% ammonium sulfate saturation and then 40-60%
saturation.
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