Mouse Reactive Necroptosis Antibody Sampler Kit #47928
Product Information
Kit Usage Information
Protocols
- 3493: Western Blotting, Immunoprecipitation (Agarose), Immunofluorescence, Flow
- 7074: Western Blotting
- 15828: Western Blotting, Immunoprecipitation (Agarose)
- 37333: Western Blotting, Immunofluorescence
- 37705: Western Blotting, Immunoprecipitation (Agarose)
- 53286: Western Blotting, Immunoprecipitation (Agarose)
- 91702: Western Blotting, Immunofluorescence
Product Description
The Mouse Reactive Necroptosis Antibody Sampler Kit provides an economical means of detecting total and phosphorylated proteins associated with necroptosis. The kit includes enough antibodies to perform two western blots with each primary antibody.
Specificity / Sensitivity
Each antibody in the Mouse Reactive Necroptosis Antibody Sampler Kit detects endogenous levels of its target protein. Phospho-RIP3 (Thr231/Ser232) (E7S1R) Rabbit mAb may not recognize RIP3 when only singly phosphorylated at either Thr231 or Ser232.
Source / Purification
Monoclonal antibodies are produced by immunizing rabbits with synthetic peptides corresponding to Leu190 of human RIP, His411 of mouse RIP3, and residues near the carboxyl terminus of mouse MLKL protein. Phospho-specific monoclonal antibodies are produced by immunizing rabbits with synthetic phosphopeptides corresponding to Ser166 of mouse RIP, Thr231/Ser232 of mouse RIP3, and Ser345 of mouse MLKL protein.
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).
In mice, activation of RIP3 is associated with phosphorylation at Thr231 and Ser232 (11), and then MLKL is phosphorylated at Ser345 by RIP3 (12).
In mice, activation of RIP3 is associated with phosphorylation at Thr231 and Ser232 (11), and then MLKL is phosphorylated at Ser345 by RIP3 (12).
- 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.
- Chen, W. et al. (2013) J Biol Chem 288, 16247-61.
- Murphy, J.M. et al. (2013) Immunity 39, 443-53.
限制使用
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For Research Use Only. Not For Use In Diagnostic Procedures.
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