Serine protease inhibitor Kazal-type 1, also called TATI or PSTI, is a protein that in humans is encoded by the SPINK1 gene. It is mapped to 5q32. SPINK1 gene contains 4 exons spanning approximately 7.5 kb. It was initially detected in urine of patients with ovarian cancer. This peptide is also produced by the mucosa of the gastrointestinal tract where it may protect the mucosal cells from proteolytic breakdown. The SPINK1 gene encodes pancreatic secretory trypsin inhibitor, which is secreted from pancreatic acinar cells into pancreatic juice. Its physiologic role was thought to be the prevention of trypsin-catalyzed premature activation of zymogens within the pancreas and the pancreatic duct. Mutations in this gene are associated with hereditary pancreatitis and tropical calcific pancreatitis. The Serine Protease Inhibitor Kazal-type 1 (SPINK1/TATI) BioAssay(TM) ELISA Kit is for the quantitative detection of human SPINK1 in cell culture supernates, serum and plasma (heparin). Detection Range: 156-10,000pg/ml Sensitivity: <10pg/ml Specificity: Natural and recombinant human SPINK1 Test Principle: This human SPINK1 BioAssay(TM) ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for SPINK1 has been precoated onto 96-well plates. Standards(E.coli, D24-C79) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for SPINK1 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 SPINK1 amount of sample captured in plate. Kit Components: 353047A: Microtiter Strips, 1x96 wells *353047B: SPINK1 Standard, 2x10ng 353047C: Anti-human SPINK1 (Biotin), 1x100ul (1:100 dilution) 143696: Avidin-Biotin-Peroxidase Complex (ABC), 1x100ul (1:100 dilution) 143697: Sample Diluent Buffer, 1x30ml 143698: Antibody Diluent Buffer, 1x12ml 143699: ABC Diluent Buffer, 1x12ml 143700: TMB Color Developing Agent, 1x10ml 143701: TMB Stop Solution, 2N H2SO4, 1x10ml 143702: Wash Buffer, 25X, 1x20ml Storage and Stability: Store *353047B powder at 4C. Once reconstituted store at 4C for up to 12 hours or at -20C for up to 48 hours. Store other components at 4C. Stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
