A versatile microassay for elastase using succinylated elastin.
We have developed a rapid, versatile, and sensitive elastase assay that is based on the measurement of primary amines that are exposed due to enzymatic degradation of proteins, using succinylated elastin as the substrate for elastase. After incubation with elastase the degree of digestion is determined with trinitrobenzene sulfonic acid. The assay is rapid and sensitive, detecting elastase down to 1 ng/ml, and is linear up to enzyme concentrations of 10 microg/ml. The assay is carried out in microtiter plate wells and therefore offers the potential for assaying numerous samples of small volume. The use of succinylated elastin shows specificity for elastase over the control protease, trypsin. This assay is also versatile because it can be applied to samples such as cell culture supernatants, blood plasma, tissue biopsies, and tissue homogenates.
Rao
S K
SK
Department of Surgery, Long Island Jewish Medical Center, Long Island Campus for Albert Einstein College of Medicine, New Hyde Park, New York 11042, USA. rao@lij.edu
Mathrubutham
M
M
Karteron
A
A
Sorensen
K
K
Cohen
J R
JR
D004796:Clinical Enzyme Tests / Q000379:methods*; D003124:Colorimetry; D004549:Elastin / Q000378:metabolism; D006801:Humans; D010196:Pancreatic Elastase / Q000097:blood / Q000378:metabolism*; D013379:Substrate Specificity; D013997:Time Factors; D014357:Trypsin / Q000378:metabolism
D004549:Elastin; D010196:Pancreatic Elastase; D014357:Trypsin