T. B. Osborne and A. J. Wakeman 245 
produced. On adding a little sodium chloride a rapidly settling 
floceulent precipitate separated, leaving very little except salt in 
the nearly clear solution. On standing a short time the pre- 
cipitate contracted to a coherent mass from which the solution 
’ was decanted almost completely. 
When this precipitate was treated with about 600 cc. of abso- 
lute alcohol and warmed it yielded a solution from which a very 
little suspended flocculent substance was removed by filtering. 
The filtrate was then concentrated 7m vacuo until much protein 
separated in consequence of the loss of alcohol. On adding ab- 
solute alcohol and warming, a syrupy solution resulted which 
remained clear on cooling. This was poured into about 3 liters 
of absolute alcohol, whereupon most of the dissolved protein was 
precipitated. On adding a few drops of a strong solution of 
ammonium acetate the rest separated, leaving the solution clear. 
The precipitate was sucked out on a Buchner funnel, washed twice 
with absolute alcohol, and three times with absolute ether. When 
dried over sulfuric acid it formed a snow-white, friable mass 
which weighed 89 gm. and was easily reduced to a fine powder. 
This was designated preparation A. 
Solution B, page 244, was evaporated to about 10 liters and 
cooled over night to room temperature. The solution was de- 
-eanted from the coherent deposit which separated. The latter 
was rinsed with water, dissolved in warm 70 per cent alcohol, its 
solution filtered clear through paper pulp, and concentrated in 
vacuo, until much of the dissolved protein separated. Absolute 
alcohol was then added and the mixture warmed. A clear syrupy 
solution resulted, which remained clear on cooling. This was 
then poured into absolute alcohol and the protein, which sepa- 
rated almost completely, was sucked out, washed with absolute 
alcohol, and then with dry ether. When dried over sulfuric acid 
46 gm. of preparation B were obtained. 
The solution from which B had first separated was further con- 
centrated to about 5 liters and, by the same procedure as em- 
ployed for B, 28 gm. of preparation C were secured. 
The 16 liters of the second alcoholic extract of the casein, 
page 244, were concentrated zn vacuo to about 3 liters and cooled 
to room temperature. Nearly all of the dissolved protein sepa- 
rated as a coherent deposit. When subjected to the same treat- 
