Description
Principle of the Assay: The Human Tryptase ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Human Tryptase in Cell Culture Supernates, Serum, Plasma. This assay employs an antibody specific for Human Tryptase coated on a 96-well plate. Standards and samples are pipetted into the wells and Tryptase present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-Human Tryptase antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of Tryptase bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
Background: Tryptases are serine proteases implicated in asthma and are highly expressed in human mast cells. They are derived from at least 4 nonallelic genes clustered on chromosome 16p13.3: TPSAB1, which represents the alpha and beta-I tryptase alleles; TPSB2, which represents the beta-II and beta-III tryptase alleles; TPSG1; and TPSD1. Elevated levels of serum tryptase occur in both anaphylactic and anaphylactoid reactions, but a negative test does not exclude anaphylaxis. Tryptase is less likely to be elevated in food allergy reactions as opposed to other causes of anaphylaxis