| Product Includes | Product # | Quantity | Color | Storage Temp |
|---|---|---|---|---|
| RIP Rabbit mAb Coated Microwells | 94588 | 96 tests |
|
+4C |
| Phospho-RIP (Ser166) Rabbit Detection mAb | 24352 | 1 ea |
|
+4C |
| HRP Diluent | 13515 | 5.5 ml |
|
+4C |
| Luminol/Enhancer Solution | 84850 | 3 ml |
|
RT |
| Stable Peroxide Buffer | 42552 | 3 ml |
|
RT |
| Sealing Tape | 54503 | 2 ea |
|
+4C |
| ELISA Wash Buffer (20X) | 9801 | 25 ml |
|
+4C |
| Cell Lysis Buffer (10X) | 9803 | 15 ml |
|
-20C |
*The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.
Description
The rapid protocol (RP) PathScan® RP Phospho-RIP (Ser166) Chemiluminescent Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of RIP protein phosphorylated at Ser166 in a reduced assay time of 1.5 hours. This chemiluminescent ELISA, which is offered in low volume microplates, shows increased signal and sensitivity while using a smaller sample size. Incubation of cell lysates and detection antibody on the coated microwell plate forms a sandwich with phospho-RIP (Ser166) protein in a single step. The plate is then extensively washed and chemiluminescent reagent is added for signal development. The magnitude of light emission, measured in relative light units (RLU), is proportional to the quantity of RIP protein phosphorylated at Ser166. Learn more about all of your ELISA kit options here.
*Antibodies in this kit are custom formulations specific to kit.
Specificity/Sensitivity
Background
The receptor-interacting protein (RIP) family of serine-threonine kinases (RIP, RIP2, RIP3, and RIP4) are important regulators of cellular stress that trigger pro-survival and inflammatory responses through the activation of NF-κB, as well as pro-apoptotic pathways (1). In addition to the kinase domain, RIP contains a death domain responsible for interaction with the death domain receptor Fas and recruitment to TNF-R1 through interaction with TRADD (2,3). RIP-deficient cells show a failure in TNF-mediated NF-κB activation, making the cells more sensitive to apoptosis (4,5). RIP also interacts with TNF-receptor-associated factors (TRAFs) and can recruit IKKs to the TNF-R1 signaling complex via interaction with NEMO, leading to IκB phosphorylation and degradation (6,7). Overexpression of RIP induces both NF-κB activation and apoptosis (2,3). Caspase-8-dependent cleavage of the RIP death domain can trigger the apoptotic activity of RIP (8).
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 (9,10). 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) (11). Research studies show that necroptosis contributes to a number of pathological conditions, and Nec-1 has been shown to provide neuroprotection in models such as ischemic brain injury (12). RIP is phosphorylated at several sites within the kinase domain that are sensitive to Nec-1, including Ser14, Ser15, Ser161, and Ser166 (13).
- Meylan, E. and Tschopp, J. (2005) Trends Biochem Sci 30, 151-9.
- Hsu, H. et al. (1996) Immunity 4, 387-96.
- Stanger, B.Z. et al. (1995) Cell 81, 513-23.
- Ting, A.T. et al. (1996) EMBO J 15, 6189-96.
- Kelliher, M.A. et al. (1998) Immunity 8, 297-303.
- Devin, A. et al. (2000) Immunity 12, 419-29.
- Zhang, S.Q. et al. (2000) Immunity 12, 301-11.
- Lin, Y. et al. (1999) Genes Dev 13, 2514-26.
- Christofferson, D.E. and Yuan, J. (2010) Curr Opin Cell Biol 22, 263-8.
- Kaczmarek, A. et al. (2013) Immunity 38, 209-23.
- Degterev, A. et al. (2008) Nat Chem Biol 4, 313-21.
- Degterev, A. et al. (2005) Nat Chem Biol 1, 112-9.
- Ofengeim, D. and Yuan, J. (2013) Nat Rev Mol Cell Biol 14, 727-36.
Background References
Cross-Reactivity Key
H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected
Trademarks and Patents
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