| 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. Sample Type: Cell lysates, cell culture supernates, serum, plasma (heparin or EDTA). Intended Use: Sandwich High Sensitivity BioAssay(TM) ELISA kit for Quantitative Detection of Human Tryptase/TPSAB1,B2. Sensitivity: <15pg/ml Range: 156pg/ml-10000pg/ml Specificity: Recognizes natural and recombinant Human Tryptase/TPSAB1,B2. There is no detectable cross-reactivity with other relevant proteins. Test Principle: This Human Tryptase/TPSAB1,B2 BioAssay(TM) ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A specific for Tryptase/TPSAB1,B2 has been precoated onto 96-well plates. Standards(Expression system for standard: NSO, Immunogen sequence: M1-V275) and test samples are added to the wells, a biotinylated detection specific for Tryptase/TPSAB1,B2 is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the Human Tryptase/TPSAB1,B2 amount of sample captured in plate. Kit Components: 96-well plate precoated with anti-Human TPSB2 antibody, 1x plate Lyophilized recombinant Human TPSB2 standard, 2x10ng/tube Biotinylated anti-Human TPSB2 antibody, 1x130ul (dilution 1:100) Avidin-Biotin-Peroxidase Complex (ABC), 1x130ul (dilution 1:100) Sample diluent buffer, 1x30ml Antibody diluent buffer, 1x12ml ABC diluent buffer, 1x12ml TMB color developing agent, 1x10ml TMB stop solution, 1x10ml PBS washing buffer Powder for 1000ml, 1x buffer Storage and Stability: Store powder at 4C liquid at -20C. Store other components at 4C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. |
