My question is: Could you explain steps? (etc. why do we heat or add chemicals like nacl or ammonium sulphate?) 1)Casein Isolation from Milk Experimental Procedure * Put 100 ml of milk in a 500 ml beaker and heat it to 40 0C. * Also heat 100 ml acetate buffer and slowly add it to the beaker with milk while stirring. * The final pH of the mixture should be 4.8. * Cool the suspension to room temperature and wait 5 more minutes before filtering. * Wash the precipitate several times with small volumes of water and then suspend it in approximately 30 ml of ethanol. Filter the suspension in the buchner funnel and wash the precipitate a second time with a mixture of equal proportions of ethanol and ether. * Finally, wash the precipitate on filter paper with 50 ml of ether and soak and dry. * Make the ether evaporate by spreading the powder. * Weigh the casein and calculate the percentage yield of the protein. * Compare with theoretical yield of 3.5 g per 100 ml milk. 2)Purification of Egg Albumin Break an egg, separate the yolk, and put the white part in a suitable beaker and add 200 ml of distilled water. Mix quickly when the albumin in the solution and globulins precipitate out. Centrifuge at 3000 g for 10 minutes to separate the supernatant and the suspension. Add the same volume of saturated ammonium sulfate solution was slowly added with stirring and let stand for 3-5 days in a cold room. Separate any albumin crystals formed.
My question is: Could you explain steps? (etc. why do we heat or add chemicals like nacl or ammonium sulphate?)
1)Casein Isolation from Milk
Experimental Procedure
* Put 100 ml of milk in a 500 ml beaker and heat it to 40 0C.
* Also heat 100 ml acetate buffer and slowly add it to the beaker with milk while stirring.
* The final pH of the mixture should be 4.8.
* Cool the suspension to room temperature and wait 5 more minutes before filtering.
* Wash the precipitate several times with small volumes of water and then suspend it in approximately 30 ml of ethanol.
Filter the suspension in the buchner funnel and wash the precipitate a second time with a mixture of equal proportions of ethanol and ether.
* Finally, wash the precipitate on filter paper with 50 ml of ether and soak and dry.
* Make the ether evaporate by spreading the powder.
* Weigh the casein and calculate the percentage yield of the protein.
* Compare with theoretical yield of 3.5 g per 100 ml milk.
2)Purification of Egg Albumin
Break an egg, separate the yolk, and put the white part in a suitable beaker and add 200 ml of distilled water.
Mix quickly when the albumin in the solution and globulins precipitate out.
Centrifuge at 3000 g for 10 minutes to separate the supernatant and the suspension.
Add the same volume of saturated ammonium sulfate solution was slowly added with stirring and let stand for 3-5 days in a cold room.
Separate any albumin crystals formed.
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