The NETosis Antibody Sampler Kit provides an economical means of detecting proteins involved in NETosis. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.
Specificity / Sensitivity
Each antibody in the NETosis Antibody Sampler Kit detects endogenous levels of its target protein. Myeloperoxidase (E1E7I) XP® Rabbit mAb recognizes the full-length and heavy chain subunits of human myeloperoxidase protein. Citrullinated Histone H3 (Arg17) (E4O3F) Rabbit mAb may also recognize histone H3 citrullinated at residue Arg26 but does not cross-react with any other known citrullinated or methylated arginine residues on histone H3. This antibody may react with a band of unknown identity at 38 kDa. Histone H3 (D1H2) XP® Rabbit mAb detects isoforms H3.1, H3.2, and H3.3. This antibody also detects the histone H3 variant CENP-A. This antibody does not cross-react with other core histones.
Source / Purification
Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Pro734 of human myeloperoxidase protein, Pro130 of human Cathepsin G protein, residues near the carboxy terminus of human neutrophil elastase protein and histone H3, and residues near the amino terminus of histone H3 in which Arg17 is citrullinated.
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 (3-5). 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 (6,7).