Description
Principle of the Assay: The Picokine™ Human PAM Pre-Coated ELISA (Enzyme-Linked Immunosorbent Assay) kit is a solid phase immunoassay specially designed to measure Human PAM with a 96-well strip plate that is pre-coated with antibody specific for PAM. The detection antibody is a biotinylated antibody specific for PAM. The capture antibody is a monoclonal antibody from mouse, the detection antibody is a biotinylated polyclonal antibody from goat. The kit contains recombinant Human PAM with immunogen: Expression system for standard: NSO, Immunogen sequence: M1-V817. The kit is analytically validated with ready to use reagents. To measure Human PAM, add standards and samples to the wells, then add the biotinylated detection antibody. Wash the wells with PBS or TBS buffer, and add Avidin-Biotin-Peroxidase Complex (ABC-HRP). Wash away the unbounded ABC-HRP with PBS or TBS buffer and add TMB. TMB is substrate to HRP and will be catalyzed to produce a blue color product, which changes into yellow after adding acidic stop solution. The density of the yellow product is linearly proportional to Human PAM in the sample. Read the density of the yellow product in each well using a plate reader, and benchmark the sample wells' readings against the standard curve to determine the concentration of Human PAM in the sample.:Background/Introduction: Peptidyl-glycine alpha-amidating monooxygenase is an enzyme that catalyzes the biosynthesis of many signaling peptides and, in humans, is encoded by the PAM gene. It is mapped to 5q21.1. This gene encodes a multifunctional protein. The encoded preproprotein is proteolytically processed to generate the mature enzyme. This enzyme includes two domains with distinct catalytic activities, a peptidylglycine alphahydroxylating monooxygenase (PHM) domain and a peptidyl-alpha-hydroxyglycine alpha-amidating lyase (PAL) domain. These catalytic domains work sequentially to catalyze the conversion of neuroendocrine peptides to active alpha-amidated products. Alternative splicing results in multiple transcript variants, at least one of which encodes an isoform that is proteolytically processed