Factors affecting the initial extraction of ovine gonadotropins.
The method of tissue homogenization as well as the solvents used for extraction influenced the protein yield, the specific activity of the oLH and oFSH, and the total units of each hormone recovered. Use of the Tekmar homogenizer (Method 2) produced a fivefold increase in the total yield of oLH per Kg glands (2390 units, Method 2; 474 units, Method 1) and the relative potency of this partially purified LH from the 80% ethanol precipitate was increased (0.16 times NIH-LH-S18, Method 2). By combining the aqueous extraction--ammonium sulfate precipitation with a subsequent ethanolic acetate extraction (Methods 3 and 4) the combined yields provided a 2.4-fold increase in oLH and 1.4-fold increase in oFSH. Thus the total units recovered of oFSH and oLH can be increased simply by changing the methods of homogenization and extraction.
D000645:Ammonium Sulfate; D000818:Animals; D001681:Biological Assay; D004591:Electrophoresis, Polyacrylamide Gel; D000431:Ethanol; D005640:Follicle Stimulating Hormone / Q000302:isolation & purification*; D007986:Luteinizing Hormone / Q000302:isolation & purification*; D008722:Methods; D010902:Pituitary Gland / Q000032:analysis*; D012756:Sheep
D000431:Ethanol; D007986:Luteinizing Hormone; D005640:Follicle Stimulating Hormone; D000645:Ammonium Sulfate