Ammonium Sulphate Precipitation Of Protein
Hello friends, this is a video lecture on precipitation of proteins, precipitation of protein by ammonium sulfate method proteins and nucleic acid play an important role in modern biology, thousands of protein and different in click acids, direct the whole cellular process for study and analysis of proteins and nucleic acid at different stage of development, this necessary to extract it. Quantitatively precipitation is widely used for the recovery of biomolecules. It is induced by using a salt or an.
Organic solvent or by changing the pH to alter the nature of solution, ammonium sulfate and sodium sulfates are the salt used for the precipitation of protein ammonium sulfate is the most widely used because it is highly soluble, inexpensive and available in the highest purity level does not change the protein solution to extreme pH. And in most of the case it does not denature proteins in native protein hydrophilic amino acids are on the surface of protein, whereas hydrophobic amino acids are buried. Attractive interaction between the nearby oppositely charged groups or ion pairs or salt bridges. It is observed that at the low concentration of salt solubility of the protein usually increases slightly. This is termed as salting in. But at high concentration of salt, the solubility of the protein drops sharply.
This is called as salting out. Now, this is the sample of protein when salt is added to an equal solution of protein, the salt require more amount of water for its dissolution. This leads to the. Computation for water molecules on the protein completely ionized, the salts have more affinity for water molecules. Then the protein. Hence, there is Salt takes ups water molecule from the protein, the ionic interaction between the water molecules and proteins are reduced.
And as a result, hydrophobic interactions dominate the hydrophobic amino acid, patches present in all the proteins, attracted each others and form aggregate. This is precipitation of proteins. This precipitation can be centrifuged. And. Collected from the tube, this precipitation can be desalted by dialysis method and further the protocol protein can be checked by any of the estimation method. Thank you.