Purification of hormone-sensitive lipase by high-performance ion exchange chromatography.
We have developed an improved method for purification of hormone-sensitive lipase from adipose tissue. The method employs two preparative high-performance ion-exchange chromatography steps on Mono Q and Mono S after detergent solubilization and partial fractionation of the enzyme by gradient sievorptive chromatography on QAE-Sephadex. About 0.2 mg of greater than 70% pure enzyme is prepared at 10% yield within 6-7 days from adipose tissue of 500 rats. This protocol is a major improvement over the previously established procedure in terms of accessibility, rapidity, enzyme purity, and yield.
D000273:Adipose Tissue / Q000201:enzymology*; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D010750:Phosphoproteins / Q000032:analysis; D051381:Rats; D002787:Sterol Esterase / Q000302:isolation & purification*
D010750:Phosphoproteins; D002787:Sterol Esterase