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  1. Products /
  2. Cellular Assay Kits /
  3. NETosis Assay Kit

NETosis Assay Kit #41855

    Product Information

    Storage

    All components in this kit are stable for at least 12 months when stored at the recommended temperature.

    Protocol

    Product Description

    The NETosis Assay Kit is a quantitative colorimetric assay that can be used to study the process of NETosis. In this kit, primary neutrophils are stimulated with PMA (included in the kit) to release neutrophil extracellular traps (NETs), leading to the production of neutrophil elastase. The unbound neutrophil elastase will be washed away, leaving behind NETs with bound neutrophil elastase. The S7 nuclease is used to digest NET DNA, releasing the neutrophil elastase from the NETs. When the neutrophil elastase substrate is added to the samples, the substrate is selectively cleaved by the neutrophil elastase to produce 4-nitroaniline that absorbs light at 405 nm. The NETosis Assay Kit does not depend upon the DNA component of NETs, as DNA release can occur independently of NETosis. The kit provides enough reagents to test up to 80 individual samples with 16 standard curve wells in a 96-well plate, or 24 samples in duplicate in a 24-well plate.

    Background

    NETosis is a unique form of regulated cell death that is characterized by membrane rupture and the extrusion of chromatin, histones, and granular and cytoplasmic components into a web-like structure called neutrophil extracellular traps (NETs) (reviewed in 1). NETosis has been associated with host defense to pathogens as well as a number of disease states, including autoimmune diseases, thrombosis, cardiovascular diseases, and tumor progression. NETosis was identified as a response to bacterial infection and can be activated by lipopolysaccharide (LPS) as well as inflammatory pathway activators like phorbol-12-myristate-13-acetate (PMA) (2). It can occur via multiple pathways, but several key players have emerged. The calcium-dependent enzyme protein-arginine deiminase 4 (PAD4) catalyzes hypercitrullination of histones that contributes to chromatin decondensation (3,4). In addition, activation of proteases, including neutrophil elastase (ELANE), myeloperoxidase (MPO), and Cathepsin G, leads to impairment of cytoskeletal structures and degradation of histones during NETosis (5,6).

    Limited Uses

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    Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

    For Research Use Only. Not For Use In Diagnostic Procedures.
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