Description
Intended Uses: For the quantitative detection of Human Chymotrypsin-like elastase family member 2A (CEL) concentration in serum, plasma and other biological fluids.
Principle of the Assay: This assay employs a two-site sandwich ELISA to quantitate CEL in Human serum, plasma. An antibody specific for CEL has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any CEL present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for CEL is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of CEL bound in the initial step. The color development is stopped and the intensity of the color is measured.
Background/Introduction: Elastases form a subfamily of serine proteases that hydrolyze many proteins in addition to elastin. Humans have six elastase genes which encode the structurally similar proteins elastase 1, 2, 2A, 2B, 3A, and 3B. Like most of the human elastases, elastase 2A is secreted from the pancreas as a zymogen. In other species, elastase 2A has been shown to preferentially cleave proteins after leucine, methionine, and phenylalanine residues. Clinical literature that describes human elastase 1 activity in the pancreas is actually referring to elastase 2A. The deduced 269-amino acid protein shares 55 to 84% identity with other mammalian elastases and 39% identity with bovine chymotrypsin A. Most homology resides in the N-terminal sequence and in the C-terminal half of the protein