PathScan® Phospho-MLKL (Ser345) Chemiluminescent Sandwich ELISA Kit #17450
Filter:
- ELISA
Supporting Data
| REACTIVITY | M |
Application Key:
- ELISA-ELISA
Species Cross-Reactivity Key:
- M-Mouse
Product Information
Product Description
The PathScan® Phospho-MLKL (Ser345) Chemiluminescent Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of MLKL protein phosphorylated at Ser345. A phospho-MLKL (Ser345) rabbit antibody has been coated onto the microwells. After incubation with cell lysates, MLKL protein phosphorylated at Ser345 is captured by the coated antibody. Following extensive washing, an MLKL mouse detection antibody is added to detect the captured phospho-MLKL (Ser345) protein. Anti-mouse IgG, HRP-linked Antibody is then used to recognize the bound detection antibody. Chemiluminescent reagent is added for signal development. The magnitude of light emission, measured in relative light units (RLU), is proportional to the quantity of phospho-MLKL (Ser345) protein.
*Antibodies in this kit are custom formulations specific to kit.
*Antibodies in this kit are custom formulations specific to kit.
Protocol
Specificity / Sensitivity
The PathScan® Phospho-MLKL (Ser345) Chemiluminescent Sandwich ELISA Kit detects endogenous levels of MLKL protein phosphorylated at Ser345. The kit sensitivity is shown in Figure 1. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
Species Reactivity:
Mouse
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), and ischemic injury (1,2). The process is negatively regulated by caspases and is initiated through a complex containing the RIP1 and RIP3 kinases, typically referred to as the necrosome. Mixed lineage kinase domain-like protein (MLKL) is a pseudokinase that was identified as a downstream target of RIP3 in the necroptosis pathway (3,4). During necroptosis RIP3 is phosphorylated at Ser227, which recruits MLKL and leads to its phosphorylation at Thr357 and Ser358 (3). Knockdown of MLKL through multiple mechanisms results in inhibition of necroptosis (3-5). While the precise mechanism for MLKL-induced necroptosis is unclear, some studies have shown that necroptosis leads to oligomerization of MLKL and translocation to the plasma membrane, where it affects membrane integrity (6-9).
- Christofferson, D.E. and Yuan, J. (2010) Curr Opin Cell Biol 22, 263-8.
- Kaczmarek, A. et al. (2013) Immunity 38, 209-23.
- Sun, L. et al. (2012) Cell 148, 213-27.
- Wang, Z. et al. (2012) Cell 148, 228-43.
- Wu, J. et al. (2013) Cell Res 23, 994-1006.
- 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.
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