A20 is a Cys2/Cys2 zinc finger protein that is induced by a variety of inflammatory stimuli and regulates gene expression. Specifically, A20 is induced by tumor necrosis factor (TNF) and interleukin 1 (IL-1), and acts as a negative regulator of nuclear factor κ B (NFκB) gene expression. By inhibiting NFκB activation, A20 plays a critical role in terminating NFκB responses to various stimuli. Although the C-terminal region of A20 contains seven zinc finger domains, only four of these domains are required for in vitro inhibition of TNF-induced NFκB activation. A20 also interacts with several other proteins, such as TRAF2, TRAF6 and IκB kinase (IKK) γ protein, and can thereby inhibit cell death. TXBP151, a novel A20-binding protein, may mediate the anti-apoptotic activity of A20. Involved in the negative feedback regulation of signal transduction, A20 and A20-binding proteins may be useful as novel therapeutic tools in the treatment of a variety of diseases.
背景文献
1. Altonsy MO et al. Context-dependent cooperation between nuclear factor B (NF-B) and the glucocorticoid receptor at a TNFAIP3 intronic enhancer: a mechanism to maintain negative feedback control of inflammation. J Biol Chem 289:8231-9 (2014).
2. Haemmig S et al. miR-125b controls apoptosis and temozolomide resistance by targeting TNFAIP3 and NKIRAS2 in glioblastomas. Cell Death Dis 5:e1279 (2014).
序列相似性
Belongs to the peptidase C64 family.
翻译后修饰
Proteolytically cleaved by MALT1 upon TCR stimulation; disrupts NF-kappa-B inhibitory function and results in increased IL-2 production. It is proposed that only a fraction of TNFAIP3 colocalized with TCR and CBM complex is cleaved, leaving the main TNFAIP3 pool intact.
Western blot analysis of TNFAIP3 on Jurkat cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1611-40, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
ICC staining of TNFAIP3 in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1611-40, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ICC staining of TNFAIP3 in A549 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1611-40, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ICC staining of TNFAIP3 in HepG2 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1611-40, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-TNFAIP3 antibody (ET1611-40) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1611-40) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Flow cytometric analysis of TNFAIP3 was done on HepG2 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1611-40, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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