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| | NETosis Assay Kit Chemical Properties |
| | NETosis Assay Kit Usage And Synthesis |
| Description | Cayman’s NETosis Assay Kit provides a simple and fast method for studying the process of NETosis ex vivo. Notably, Cayman’s NETosis Assay Kit does not depend upon the DNA component of NETs, as DNA release can occur independently of NETosis. In this kit, primary neutrophils are stimulated to release NETs with either PMA or a calcium ionophore (both included). As shown in Figure 1, unbound neutrophil elastase is washed away following NET generation. Following digestion of NET DNA by S7 nuclease, the supernatant containing neutrophil elastase is added to a substrate, which is selectively cleaved by neutrophil elastase to yield a 4-nitroaniline product that absorbs light at 405 nm. A sufficient amount of each reagent is provided to test and analyze up to 80 individual samples or 24 samples in duplicate.Needed but not supplied: Please download the kit booklet to verify if UltraPure Water (Milli-Q or equivalent) or any other components are needed for this assay.WARNING This product is not for human or veterinary use. | | 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. 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 as well as inflammatory pathway activators like phorbol-12-myristate-13-acetate. It can occur via multiple pathways, but several key players have emerged. The calcium-dependent enzyme protein-arginine deiminase 4 catalyzes hypercitrullination of histones that contributes to chromatin decondensation. In addition, activation of proteases, including neutrophil elastase, myeloperoxidase, and Cathepsin G, leads to impairment of cytoskeletal structures and degradation of histones during NETosis. | | References | [1] Thiam, H.R. et al. (2020) Annu Rev Cell Dev Biol 36, 191-218.
[2] Brinkmann, V. et al. (2004) Science 303, 1532-5.
[3] Li, P. et al. (2010) J Exp Med 207, 1853-62.
[4] Wong, S.L. and Wagner, D.D. (2018) FASEB J, fj201800691R.
[5] Papayannopoulos, V. et al. (2010) J Cell Biol 191, 677-91.
[6] Metzler, K.D. et al. (2014) Cell Rep 8, 883-96. |
| | NETosis Assay Kit Preparation Products And Raw materials |
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