Elastase/ELANE/ELA2 Picokine (ELANE) ELISA Kit for Mouse

Principle of the Assay: The Picokine™ Mouse Elane Pre-Coated ELISA (Enzyme-Linked Immunosorbent Assay) kit is a solid-phase immunoassay specially designed to measure Mouse Elane with a 96-well strip plate that is pre-coated with antibody specific for Elane. The detection antibody is a biotinylated antibody specific for Elane. The capture antibody is a monoclonal antibody from rat and the detection antibody is a biotinylated polyclonal antibody from goat. The kit contains recombinant Mouse Elane with immunogen: Expression system for standard: NS0; Immunogen sequence: S27-R260. The kit is analytically validated with ready-to-use reagents. To measure Mouse Elane, 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 an HRP substrate and will be catalyzed to produce a blue color product, which changes into yellow after adding the acidic stop solution. The absorbance of the yellow product at 450nm is linearly proportional to Mouse Elane in the sample. Read the absorbance 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 Mouse Elane in the sample.

Background/Introduction: Neutrophil elastase is a serine protease of neutrophil and monocyte granules. Its key physiologic role is in innate host defense, but it can also participate in tissue remodeling and possesses secretagogue actions important to local inflammatory responses. Elastases form a subfamily of serine proteases that hydrolyze many proteins in addition to elastin. Humans have six elastase genes which encode structurally similar proteins. The encoded preproprotein is proteolytically processed to generate the active protease. Following activation, this protease hydrolyzes proteins within specialized neutrophil lysosomes, called azurophil granules, as well as proteins of the extracellular matrix. This protein also degrades the outer membrane protein A (OmpA) of E. coli as well as the virulence factors of such bacteria as Shigella, Salmonella and Yersinia. Mutations in this gene are associated with cyclic neutropenia and severe congenital neutropenia (SCN). This gene is present in a gene cluster on chromosome 19.

Small volumes of Elane elisa kit vial(s) may occasionally become entrapped in the seal of the product vial during shipment and storage. If necessary, briefly centrifuge the vial on a tabletop centrifuge to dislodge any liquid in the container`s cap. Certain products may require to ship with dry ice and additional dry ice fee may apply.

MBS177639 is a ready-to-use microwell, strip plate ELISA (enzyme-linked immunosorbent assay) Kit for analyzing the presence of the Elastase/ELANE/ELA2 PicoKine, ELISA Kit target analytes in biological samples. The concentration gradients of the kit standards or positive controls render a theoretical kit detection range in biological research samples containing Elane. The ELISA analytical biochemical technique of the MBS177639 kit is based on Elane antibody-Elane antigen interactions (immunosorbency) and an HRP colorimetric detection system to detect Elane antigen targets in samples. The ELISA Kit is designed to detect native, not recombinant, Elane. Appropriate sample types may include undiluted body fluids and/or tissue homogenates, secretions. Quality control assays assessing reproducibility identified the intra-assay CV (%) and inter-assay CV(%).