A protein is a naturally occurring, unbranched polymer in which the monomers
units are amino acids, which at least has 50 amino acids residues. The arrangement of these amino acids contributes in the unique physico-chemical characteristics of a protein which determine what protein isolation and purification method should be done. To denature a protein is to break its native conformation by subjecting the protein to extreme heat and pH, introducing chaotropic agents, or infusing it with chemicals that can break disulfide bonds. To isolate casein from skimmed milk, an acid is used to adjust the pH to 4.6 which is the isoelectric pH of the casein. The isolated casein has undergone acid hydrolysis; this process is done to obtain information about the isolated caseins composition, then, protein is treated with an acid, alkali, or proteolytic enzymes. For Qualitative color reactions, after auto- claving, the intact proteins were used in different tests such as Biuret test, Ninhydrin test , Xanthoproteic test , Millons test, Hopkins-Cole test, Sakaguchi test, Nitroprusside test, Fohls, test for amides, and Paulys test. Paper chromatography was performed for the separation and identification of amino acid standards based on the polarities of tryptophan, arginine, proline, cysteine, serine, aspartate, histidine, glycine, and alanine.