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Render Timestamp: 2024-07-27T18:37:47.380Z
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PDP - Template Name: Monoclonal Antibody
PDP - Template ID: *******c5e4b77
R Recombinant
Recombinant: Superior lot-to-lot consistency, continuous supply, and animal-free manufacturing.

Phospho-RIP (Ser321) (E9K2A) Rabbit mAb (BSA and Azide Free) #33527

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    Supporting Data

    REACTIVITY M
    SENSITIVITY Endogenous
    MW (kDa) 78
    Source/Isotype Rabbit IgG
    Application Key:
    • WB-Western Blotting 
    • IF-Immunofluorescence 
    • F-Flow Cytometry 
    Species Cross-Reactivity Key:
    • M-Mouse 

    Product Information

    Product Usage Information

    This product is the carrier free version of product #38662. All data were generated using the same antibody clone in the standard formulation which contains BSA and glycerol.

    This formulation is ideal for use with technologies requiring specialized or custom antibody labeling, including fluorophores, metals, lanthanides, and oligonucleotides. It is not recommended for ChIP, ChIP-seq, CUT&RUN or CUT&Tag assays. If you require a carrier free formulation for chromatin profiling, please contact us. Optimal dilutions/concentrations should be determined by the end user.

    Formulation

    Supplied in 1X PBS, BSA and Azide Free.

    For standard formulation of this product see product #38662

    Storage

    Store at -20°C. This product will freeze at -20°C so it is recommended to aliquot into single-use vials to avoid multiple freeze/thaw cycles. A slight precipitate may be present and can be dissolved by gently vortexing. This will not interfere with antibody performance.

    Specificity / Sensitivity

    Phospho-RIP (Ser321) (E9K2A) Rabbit mAb (BSA and Azide Free) recognizes endogenous levels of mouse RIP protein only when phosphorylated at Ser321.


    Species Reactivity:

    Mouse

    Source / Purification

    Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser321 of mouse RIP protein.

    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).

    RIP is also phosphorylated at Ser321(mouse)/Ser320(human) by MAPKAPK-2 (MK-2) and TAK1 in response to inflammatory signals such as TNF-α and LPS (14-17). Phosphorylation at this site suppresses RIP mediated apoptosis by inhibiting its interaction with FADD and caspase-8 (14-17).

    1. Meylan, E. and Tschopp, J. (2005) Trends Biochem Sci 30, 151-9.
    2. Hsu, H. et al. (1996) Immunity 4, 387-96.
    3. Stanger, B.Z. et al. (1995) Cell 81, 513-23.
    4. Ting, A.T. et al. (1996) EMBO J 15, 6189-96.
    5. Kelliher, M.A. et al. (1998) Immunity 8, 297-303.
    6. Devin, A. et al. (2000) Immunity 12, 419-29.
    7. Zhang, S.Q. et al. (2000) Immunity 12, 301-11.
    8. Lin, Y. et al. (1999) Genes Dev 13, 2514-26.
    9. Christofferson, D.E. and Yuan, J. (2010) Curr Opin Cell Biol 22, 263-8.
    10. Kaczmarek, A. et al. (2013) Immunity 38, 209-23.
    11. Degterev, A. et al. (2008) Nat Chem Biol 4, 313-21.
    12. Degterev, A. et al. (2005) Nat Chem Biol 1, 112-9.
    13. Ofengeim, D. and Yuan, J. (2013) Nat Rev Mol Cell Biol 14, 727-36.
    14. Jaco, I. et al. (2017) Mol Cell 66, 698-710.e5.
    15. Geng, J. et al. (2017) Nat Commun 8, 359.
    16. Dondelinger, Y. et al. (2017) Nat Cell Biol 19, 1237-47.
    17. Menon, M.B. et al. (2017) Nat Cell Biol 19, 1248-59.
    For Research Use Only. Not For Use In Diagnostic Procedures.
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