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.
Sample Type:Cell culture supernates, serum and plasma (heparin)
Intended Use:For quantitative detection of human SPINK1 in cell culture supernates, serum and plasma(heparin).
Sensitivity:<10pg/ml
Range:156-10,000pg/ml
Specificity:Natural and recombinant human SPINK1
Test Principle:This human SPINK1 BioAssay™ 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:96-well plate precoated with human SPINK1 antibody, 1Lyophilized recombinant human SPINK1 standard, 10ng/tube×2Biotinylated human SPINK1 antibody, 130ul (dilution 1:100)Avidin-Biotin-Peroxidase Complex (ABC), 130ul (dilution 1:100)Sample diluent buffer, 30mlAntibody diluent buffer, 12mlABC diluent buffer, 12mlTMB color developing agent, 10mlTMB stop solution, 10ml
Storage and Stability:Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
