Necroptosis Antibody Sampler Kit #98110
Product Information
Kit Usage Information
Protocols
- 3493: Western Blotting, Immunoprecipitation (Agarose), Immunofluorescence, Flow
- 7074: Western Blotting
- 13526: Western Blotting, Immunoprecipitation (Agarose)
- 14993: Western Blotting
- 65746: Western Blotting
- 91689: Western Blotting
- 93654: Western Blotting, Immunofluorescence
Product Description
Specificity / Sensitivity
Each antibody in the Necroptosis Antibody Sampler Kit detects endogenous levels of its target protein. MLKL (D2I6N) Rabbit mAb cross-reacts with an unidentified band at 130 kDa in some cell lines. Phospho-MLKL (Ser358) (D6H3V) Rabbit mAb may also bind to MLKL when dually phosphorylated at Thr357 and Ser358.
Source / Purification
Monoclonal antibodies are produced by immunizing rabbits with synthetic peptides corresponding to Leu190 of human RIP, residues near the carboxyl terminus of human RIP3, and residues near the carboxyl terminus of human MLKL. Phospho-specific monoclonal antibodies are produced by immunizing rabbits with synthetic phospho-peptides corresponding to Ser166 of human RIP, Ser227 of human RIP3, and Ser358 of human MLKL.
Background
Necroptosis, a regulated pathway for necrotic cell death, is triggered by a number of inflammatory signals, including cytokines in the tumor necrosis factor (TNF) family, pathogen sensors such as toll-like receptors (TLRs), ischemic injury, and neurodegenerative diseases (1-3). The process is negatively regulated by caspases and is initiated through a complex containing the RIP and RIP3 kinases, typically referred to as the necrosome. Necroptosis is inhibited by a small molecule inhibitor of RIP, necrostatin-1 (Nec-1) (4). RIP is phosphorylated at several sites within the kinase domain that are sensitive to Nec-1, including Ser14, Ser15, Ser161, and Ser166 (5). During necroptosis, RIP3 is phosphorylated at Ser227, leading to recruitment and phosphorylation of MLKL at Thr357 and Ser358 (6). Phosphorylation of MLKL results in its oligomerization and translocation to the plasma membrane, where it affects membrane integrity (7-10).
- Christofferson, D.E. and Yuan, J. (2010) Curr Opin Cell Biol 22, 263-8.
- Kaczmarek, A. et al. (2013) Immunity 38, 209-23.
- Zhou, W. and Yuan, J. (2014) Semin Cell Dev Biol 35, 14-23.
- Degterev, A. et al. (2008) Nat Chem Biol 4, 313-21.
- Ofengeim, D. and Yuan, J. (2013) Nat Rev Mol Cell Biol 14, 727-36.
- Sun, L. et al. (2012) Cell 148, 213-27.
- Cai, Z. et al. (2014) Nat Cell Biol 16, 55-65.
- Chen, X. et al. (2014) Cell Res 24, 105-21.
- Wang, H. et al. (2014) Mol Cell 54, 133-46.
- Dondelinger, Y. et al. (2014) Cell Rep 7, 971-81.
Limited Uses
Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.
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.