Bay11 7082

Manufactured by Apexbio

Sourced in United States

BAY11-7082 is a laboratory reagent used for research purposes. It is an inhibitor of the NF-κB signaling pathway. The core function of BAY11-7082 is to inhibit the phosphorylation of IκBα, which is a key regulatory step in the activation of NF-κB.

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5 protocols using bay11 7082

Cell Viability Assay with Celecoxib, AH6809, and BAY11-7082

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Cell viability was measured using a Cell Counting Kit-8 (CCK-8) assay (Dojindo Laboratories, Kumamoto, Japan). Cells were seeded in a 96-well flat-bottomed plate (2 × 10³ cells in 100 μl per well), incubated overnight to allow cell attachment, and exposed to 100 mM celecoxib (Pfizer, New York, NY, USA), 10 μM AH6809 (ApexBio Technology, Houston, TX, USA) or 10 μM BAY11-7082 (ApexBio Technology) for 24, 48, 72, and 96 hour. Then 10 µl of CCK-8 solution was added to each well, the cells were incubated for another 2 hours, and the absorbance at 450 nm was measured with a microplate reader. All experiments were performed thrice.

Zhang X., Yan K., Deng L., Liang J., Liang H., Feng D, & Ling B. (2019). Cyclooxygenase 2 Promotes Proliferation and Invasion in Ovarian Cancer Cells via the PGE2/NF-κB Pathway. Cell Transplantation, 28(1 Suppl), 1S-13S.

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Acute Inflammation Response in Mice

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The littermate male WT and Lf^−/− mice (6–8 weeks) were weighed and injected intraperitoneally with LPS (L9143, Sigma–Aldrich) at a dose of 4 mg/kg to induce acute inflammation response (or with an equal volume of PBS as the control group, each group n = 5). For Cenicriviroc treatment experiment, the mice were intraperitoneally injected with Cenicriviroc (CVC, 10 mg/kg, Selleck) 10 min prior to LPS injection. 4 h later, the mice were weighed again (each group n = 3). Serum was collected and stored at − 80 °C. The mice were injected intraperitoneally with 5 ml of RPMI-1640 after euthanasia, and the peritoneal lavage fluid was collected and centrifuged to obtain the lavage fluid supernatant and peritoneal cells. The WT and Lf^−/− MEF cells were stimulated by LPS (1 μg/ml) or PBS for 4 h, and BAY11-7082 (1 μM, APExBIO) is added to inhibit the NF-κB pathway.

Liu C., Peng Q., Wei L., Li Z., Zhang X., Wu Y., Wang J., Zheng X., Wen Y., Zheng R., Yan Q., Ye Q, & Ma J. (2022). Deficiency of Lactoferrin aggravates lipopolysaccharide-induced acute inflammation via recruitment macrophage in mice. Biometals, 36(3), 549-562.

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Solubilization of Oncological Compounds

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A stock solution of imatinib (Sigma-Aldrich) was dissolved in water, and decitabine (Sigma-Aldrich) and BAY11-7082 (APExBIO) were dissolved in dimethyl sulfoxide (DMSO). All stock solutions were stored at -20°C.

Wang X., Hou Y., Lyu Y., Zhou J., Zhang X., Hassani M.A., Huang D., Zhao Z., Zhou D., Xie F., Zhang X, & Yan J. (2024). LncRNA IRAIN overcomes imatinib resistance in chronic myeloid leukemia via NF-κB/CD44 pathway inhibition. iScience, 27(6), 109851.

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Modulation of Cell Signaling Pathways

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PC12 or C17.2 cells were seeded in a 6-well plate at the density of 1 × 10⁶ cells/well and incubated for 48 h until reaching 80 % confluence. Then, cells were treated with 10% CM (v/v) from M. smegmatis, or 0.5 ng/ml interferon gamma (IFN-γ) (ab645, abcam, USA), or anti-INFγ (1:1,000 dilution, AF-585-NA, R&D system, USA) for 48 h. For positive control, PC12 cells were treated with 50 ng/ml nerve growth factor (NGF Sigma, USA), and C17.2 cells were treated with 50 ng/ml NGF and 50 ng/ml brain-derived neurotrophic factor (BDNF, Sigma, USA. For screening experiment of signal pathway, cells were treated with CM and different inhibitors (AG490, SP600125, U0126, SB239063, PD98059, H89, Y27632, IWP2, DAPT, BAY 11-7082, LY294002, APExBIO Technology, Houston, USA) for 48 h respectively. The morphology of the cells was observed by microscope (Nikon TE 300).

Feng X., Lu J., He Z., Wang Y., Qi F., Pi R, & Zhang G. (2018). Mycobacterium smegmatis Induces Neurite Outgrowth and Differentiation in an Autophagy-Independent Manner in PC12 and C17.2 Cells. Frontiers in Cellular and Infection Microbiology, 8, 201.

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Prostate Cancer Cell Lines Characterization

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LNCaP, PC3, DU145, C4-2B cell lines were originally acquired from ATCC. 22Rv1 cell line was provided by Stem Cell Bank, Chinese Academy of Sciences. VCaP WT and VCaP ENZR were kindly provided from Lab Zhenfei Li, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences. LNCaP, PC3, DU145, C4-2B, and 22Rv1 were cultured in RPMI 1640 (HyClone) with 10% fetal bovine serum (FBS; Gibco). 293 T and VCaP were cultured in DMEM/High Glucose (HyClone) with 10% FBS at 37 °C with 5% CO₂. All cell lines were tested mycoplasma negative using DAPI staining. Enzalutamide, Z-Vad-FMK, Bay 11–7082, and Reparixin inhibitors were purchased from ApexBio. Enzalutamide (100 mM), Z-Vad-FMK (50 mM), Bay 11–7082(50 mM), and Reparixin (40 mM) were suspended in DMSO. hTNF-α (200 μg/ml) was purchased from Biolegend. All antibodies used in were list in Table S3.

Xia J., Zhang J., Wang L., Liu H., Wang J., Liu J., Liu Z., Zhu Y., Xu Y., Yang W, & Yu Y. (2021). Non-apoptotic function of caspase-8 confers prostate cancer enzalutamide resistance via NF-κB activation. Cell Death & Disease, 12(9), 833.

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