Pure casein salt solutions and fresh milk do not coagulate on boiling, but in the presence of free acid coagulation may take place below the boiling temperature. The coagulum formed in the case of milk includes fat and calcium phosphate. The slight pellicle which coats over milk when it is warmed is of the same composition. +49. Use of acid.+--A commonly accepted explanation of the precipitation of casein by acids is that the casein is held in solution by chemical union with a base (lime in the case of milk); that added acid removes the base, allowing the insoluble casein to precipitate; and that excess of acid unites with casein, forming a compound which is more or less readily soluble. +50. Robertson's theory.+--According to Robertson's conception, in a soluble solution of a protein or its salt, the molecules of the protein unite with each other to a certain extent, in this way forming polymers. The reaction is reversible, and the point of equilibrium between the compound and its polymeric modification varies under the influence of whatever condition affects the concentration of the protein ions. Addition of water, or of acid, alkali or salt, or the application of heat has such an effect, and consequently alters the relative number of heavier molecule-complexes. Robertson's experiments give evidence that one of the effects of increase of temperature on a solution of casein is a shifting of the equilibrium in the direction of the higher complexes. He explains coagulation as being a result of these molecular aggregates becoming so large as to assume the properties of matter in mass and to become practically an unstable suspension and then a precipitate. The acid curd then is casein or some combination of casein with the precipitant acid. +51. Rennet curd.+--Rennet extract and pepsin coagulation differs from coagulation by acids, and cannot be looked on as a simple removal of the base from a caseinate. The presence of soluble calcium salts (or other alkaline earth salts) seems to be essential, and the precipitate formed is not casein or a casein salt, but a salt of a slightly different nucleoalbumin called "paracasein." Many writers, following Halliburton, call this modification produced by rennin the "casein" and that from which it is derived, "caseinogen." Foster and a few others have used the term "tyrein" for the rennet clot. A number of investigations have been made on the conditions essential or favorable to formation of the coagulum, especially with regard to the effects of the degree of acidity and of conditions affecting the amount of calcium present, either as free soluble salt or bound to the casein. Soluble salts of calcium, barium and strontium favor or hasten coagulation, while salts of ammonium, sodium and potassium retard or prevent coagulation.