Bay 11 7082

Manufactured by Bio-Techne

Sourced in United Kingdom, United States

BAY 11-7082 is a chemical compound that functions as an inhibitor of IκBα phosphorylation. It is commonly used in research applications to study the NF-κB signaling pathway.

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Lab products found in correlation

Ly294002, by Bio-Techne (4 mentions) Pd98059, by Bio-Techne (3 mentions) Dimethyl sulfoxide (dmso), by Merck Group (2 mentions) Lipopolysaccharides (lps), by Merck Group (2 mentions) Pam3csk4, by InvivoGen (2 mentions) Sp600125, by Bio-Techne (2 mentions) Nf κb p65, by Santa Cruz Biotechnology (2 mentions) P38 mapk, by Santa Cruz Biotechnology (2 mentions) β actin, by Santa Cruz Biotechnology (2 mentions) Rpmi 1640, by Thermo Fisher Scientific (1 mentions) Sodium pyruvate, by Thermo Fisher Scientific (1 mentions) Glutamax, by Thermo Fisher Scientific (1 mentions) Lipopolysaccharides (lps), by InvivoGen (1 mentions) Flagellin, by InvivoGen (1 mentions) Rnaprotect cell reagent, by Qiagen (1 mentions) Gentamicin, by Lonza (1 mentions) A438079, by Bio-Techne (1 mentions) Rpmi 1640 culture medium, by Thermo Fisher Scientific (1 mentions) Mtt reagent, by Merck Group (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)

14 protocols using bay 11 7082

Monocyte Stimulation Experiments

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For stimulation experiments of CD14⁺ monocytes, cells were seeded in a 48-well plate (677970; Greiner Bio-One, Frickenhausen, Germany) at a density of 5 × 10⁵ cells per well in 400 µl of RPMI 1640 (Gibco) with 10% sterile FCS, gentamicin (10 µg/ml; Lonza Bioscience, Durham, NC), 2 mM GlutaMAX (Thermo Fisher Scientific, Waltham, MA), and 1 mM sodium pyruvate (Thermo Fisher Scientific, Waltham, MA). Cells were subsequently stimulated with medium control, LPS (from Escherichia coli O111:B4; 100 ng/ml; In vivo gen, San Diego, CA), Pam3CSK4 (1 µg/ml; In vivo gen), flagellin (from P. aeruginosa; 1 µg/ml; FLA-PA Ultrapure, In vivo gen), and poly(I:C) High Molecular Weight (HMW) (10 µg/ml, In vivo gen) for 2 h or heat-killed bacteria [cell-bacteria ratio, 1:10; Streptococcus (S.) pneumoniae ATCC^® 6303 and Klebsiella (K.) pneumoniae ATCC43816] for 2, 6, and 24 h. Medium was collected, and the cells were stored in RNA protect cell reagent (Qiagen, Hilden, Germany) until further processing. In a separate experiment, primary monocytes (5 × 10⁵cells per well) were pretreated with the NF-κB inhibitor BAY11-7082 (Malladi et al., 2004 (link)) (10 µM; Tocris Bioscience, Bristol, UK) or vehicle control (dimethyl sulfoxide; Merck, Darmstadt, Germany) for 30 min and then incubated in medium, with or without LPS (100 ng/ml) for 2 h.

Malmström E., Khan H.N., Veer C.V., Stunnenberg M., Meijer M.T., Matsumoto H., Otto N.A., Geijtenbeek T.B., de Vos A.F., van der Poll T, & Scicluna B.P. (2022). The Long Non-Coding Antisense RNA JHDM1D-AS1 Regulates Inflammatory Responses in Human Monocytes. Frontiers in Cellular and Infection Microbiology, 12, 934313.

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Spinal Cord Injury Mouse Model

Cited 2 times

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Mice were anesthetized with ketamine (75 mg/kg) and xylazine (3 mg/kg) intraperitoneally and the muscles overlying the T5 to T8 vertebrae were dissected, followed by laminectomy to expose the spinal cord. To induce SCI, extradural compression of the spinal cord at the level of the T6-T7 vertebra was conducted via placement of an aneurysm clip with a closing force for 1 min, as described by Paterniti et al. [17 (link)]. The aneurysm clip is with a closing force of 30 g based on previous study [18 (link)]. Mice undergoing laminectomy alone served as the sham group.
Mice in treatment groups received a daily single-dose intraperitoneal injection of BAY 11-7082 (Tocris Bioscience, Bristol, UK) at 20 mg/kg or A438079 (Tocris Bioscience) at 80 mg/kg in 0.1 ml vehicle (DMSO and 0.9% NaCl, 1:3) immediately after injury and following 2 days. Sham and SCI groups received daily 0.1 ml vehicle immediately post injury and following 2 days intraperitoneally. Mice underwent manual bladder expression twice a day until recovery of reflex bladder emptying. The timing and dose of BAY 11-7082 [19 (link), 20 (link)] and A438079 [21 (link)] were based on previous studies.

Jiang W., Li M., He F., Zhou S, & Zhu L. (2017). Targeting the NLRP3 inflammasome to attenuate spinal cord injury in mice. Journal of Neuroinflammation, 14, 207.

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Lung Slice Culture Protocol for In Vitro Studies

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PCLS of 300 μm were generated from the lungs of 3-month-old β₁^AT2-KO and β₁^fl/fl mice, as previously described (56 (link)–58 (link)). Briefly, slices were cultured in DMEM:F12 for 12 hours, then incubated with BrdU (1 mM, MilliporeSigma B5002) for 4 hours prior to 48 hours of treatment with LPS (MilliporeSigma L2880, 62.5 ng/mL) and/or BAY 11-7082 (100 μM, Tocris, Bio-Techne; Bay11-7821). Slices were then fixed, paraffin-embedded, and sectioned.

Sucre J.M., Bock F., Negretti N.M., Benjamin J.T., Gulleman P.M., Dong X., Ferguson K.T., Jetter C.S., Han W., Liu Y., Kook S., Gokey J.J., Guttentag S.H., Kropski J.A., Blackwell T.S., Zent R, & Plosa E.J. (2023). Alveolar repair following LPS-induced injury requires cell-ECM interactions. JCI Insight, 8(14), e167211.

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Molecular Immunology of Anti-Inflammatory Agents

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Penicillin-streptomycin antibiotic, FBS, PBS, and RPMI-1640 culture medium were attained from Gibco (Grand Island, NY, USA). R & D Systems (Minneapolis, MN, USA) were the supplier of ELISA kits for human IL-1β, TNF-α and PGE₂. MTT reagent, LPS, RIPA, and DMSO were obtained from Sigma Chemical Co. (St. Louis, MO, USA). Levamisole (purity > 98%) was procured from Cayman Chemical (Ann Arbor, MI, USA). Tocris Biosciences (Bristol, UK) supplied Akt inhibitor (LY294002), p38 inhibitor (SB202190), ERK inhibitor (U0126), JNK inhibitor (SP600125), and NF-κB inhibitor (BAY 11–7082). Pierce (Rockford, IL, USA) supplied 1 × Halt phosphatase and protease inhibitor cocktail. Cell Signaling Technology (Beverly, MA) supplied primary antibodies specific to p-NFκBp65, p-IKKα/β, p-JNK, IκBα, p-IκBα, p-ERK, p-p38, JNK, ERK, p38, COX-2, p-Akt, TRL4 and MyD88 along with anti-rabbit secondary antibody conjugated to horseradish peroxidase and β-actin.

Haque M.A., Jantan I., Harikrishnan H, & Ahmad W. (2020). Standardized ethanol extract of Tinospora crispa upregulates pro-inflammatory mediators release in LPS-primed U937 human macrophages through stimulation of MAPK, NF-κB and PI3K-Akt signaling networks. BMC Complementary Medicine and Therapies, 20, 245.

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Isolation and Stimulation of Monocytes

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Buffy coats were collected from heparin anticoagulated blood by density-gradient centrifugation using Ficoll-paque plus (GE Healthcare, Stockholm, Sweden). Subsequently, monocytes were isolated using cluster-of-differentiation (CD)14⁺ magnetic beads (Miltenyi Biotec, Bergisch Gladbach, Germany, # 130-050-201) and MACS LS columns (Miltenyl Biotech). For stimulations, monocytes were placed in a 48-well plate (10⁶ per well; Greiner Bio-One, Frickenhausen, Germany) in RPMI1640 medium containing 10% fetal calf serum (FCS),100 U/ml penicillin, 2 mM L-glutamine and stimulated with medium, Pam3CSK4 (1 µg/mL; Invivogen, San Diego, CA) or LPS (from Escherichia coli O111:B4;100 ng/mL; Invivogen) for 4 hours. In a separate experiment, monocytes (5 × 10⁵ cells/well) were pretreated with the NF-κB inhibitor BAY11-7082 (10 µM; Tocris Bioscience, Bristol, UK) or vehicle control (dimethyl sulfoxide; Merck KGaA, Darmstadt, Germany) for 30 minutes, and then incubated in medium, with or without LPS (100 ng/mL) for 2 hours.

Matsumoto H., Scicluna B.P., Jim K.K., Falahi F., Qin W., Gürkan B., Malmström E., Meijer M.T., Butler J.M., Khan H.N., Takagi T., Ishii S., Schultz M.J., van de Beek D., de Vos A.F., van ‘t Veer C, & van der Poll T. (2021). HIVEP1 Is a Negative Regulator of NF-κB That Inhibits Systemic Inflammation in Sepsis. Frontiers in Immunology, 12, 744358.

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Investigating Cytokine Signaling in Human Mesangial Cells

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Human mesangial cells (HMC)48 (link) were grown in DMEM containing 10% FCS, 100 units/ml penicillin, and 100 mg/ml streptomycin. Recombinant human IL-1α, IL-1β, IL-1Ra, human CXCL1, 2, 8 ELISA kits were purchased from PeproTech (Rocky Hill, NJ, USA). Mouse anti-human IL-1R1 antibody was purchased from Abcam (Cambridge, UK). Rabbit anti-human phosphorylated ERK, JNK, and p38 antibodies, rabbit anti-human A20 antibody, and ERK inhibitor U0126 were purchased from Cell Signaling Technology (Beverly, MA, USA). P38 inhibitor SB203580, JNK inhibitor SP600125, and NF-κB inhibitor Bay117082 were purchased from Tocris (Ellisville, MO, USA). A20 mammalian expressing plasmid (EX-K6040-M11) was purchased from Genecopoeia (Rockville, MD, USA).

Luo H., Liu Y., Li Q., Liao L., Sun R., Liu X., Jiang M, & Hu J. (2015). A20 regulates IL-1-induced tolerant production of CXC chemokines in human mesangial cells via inhibition of MAPK signaling. Scientific Reports, 5, 18007.

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Plasmacytoid Dendritic Cell Stimulation

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RPMI 1640 (Biochrom) supplemented with 10% heat-inactivated FCS (Invitrogen), 1 mM L-glutamine, 100 U/mL penicillin, 100 μg/mL streptomycin (all Sigma-Aldrich), and 10 ng/mL IL-3 (Peprotech) or 10 ng/mL GM-CSF (ImmunoTools) was used. Recombinant human IFN-β was from Peprotech. Completely and partially phosphorothioate-modified ODNs were from Metabion. Small letters indicate phosphorothioate linkage, whereas capital letters stand for phosphodiester linkage 3′ of the base: ODN 2006: 5′-tcgtcgttttgtcgttttgtcgtt-3′; ODN 2216: 5′-ggGGGACGATCGTCgggggG-3′. R848 was from Invivogen. The single-stranded RNA oligonucleotide RNA9.2sense: 5′-AGCUUAACCUGUCCUUCAA-3′ [38 ] was from Eurogentec. ssRNA and DNA oligonucleotides were complexed with poly-l-arginine as previously described [39 ]. For blocking experiments, the neutralizing anti-IFN-receptor chain 2 antibody (PBL Biomedical Laboratories) was employed. pDCs were preincubated with this antibody or a respective isotype control (mouse IgG2a) at 10 μg/mL for 30 min prior to stimulation. To inhibit protein synthesis, pDCs were treated with cycloheximide (Sigma-Aldrich) 30 min prior to stimulation. Bay 11–7082 and SR 11302 were from Tocris Bioscience. The IFN-α ELISA was from Bender MedSystems.

Kim S., Kaiser V., Beier E., Bechheim M., Guenthner-Biller M., Ablasser A., Berger M., Endres S., Hartmann G, & Hornung V. (2014). Self-priming determines high type I IFN production by plasmacytoid dendritic cells. European journal of immunology, 44(3), 807-818.

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Isolation and Characterization of Human Dendritic Cells

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RPMI 1640, fetal bovine serum (FBS), penicillin/streptomycin and L-glutamine were purchased from Euroclone, Devon, UK. Fycoll was obtained from Cederlane Labs and Percoll from Amersham Bioscience, Pittsburgh, PA, USA. Recombinant human granulocyte macrophage colony stimulating factor (GM-CSF) and interleukin-13 (IL-13) were purchased from ProSpec TechnoGene, East Brunswick, NJ, USA. All the reagents contained <0.125 endotoxin units/mL, as checked by the Limulus amebocyte lysate assay (Cambrex, East Rutherford, NJ, USA). LPS from Escherichia coli strain 026:B6 was obtained from Sigma-Aldrich, Milano, Italy. Wortmannin, LY 294002, U0126 and BAY 11-7082 were purchased from Tocris Biosciences, Bristol, UK. SAR405 was purchased from CliniSciences, Nanterre, France. Baf A1 was obtained Enzo Life Sciences, Plymouth Meeting, PA, USA.

Monaci S., Coppola F., Rossi D., Giuntini G., Filippi I., Marotta G., Sozzani S., Carraro F, & Naldini A. (2022). Hypoxia Induces Autophagy in Human Dendritic Cells: Involvement of Class III PI3K/Vps34. Cells, 11(10), 1695.

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Inhibitor Compounds for Cell Studies

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N-p-tosyl-L-phenylalanine chloromethyl ketone (TPCK, # T4376), N-α-tosyl-L-lysine chloromethyl ketone hydrochloride (TLCK, #90182), suberoylanilide hydroxamic acid (SAHA, #SML0061), anacardic acid (#A7236), decitabine (#A3656), lipopolysaccharide (LPS from Escherichia coli O111:B4, #L4391) were purchased from Sigma-Aldrich. Carfilzomib (#S2853) and tocilizumab (#A2012) were purchased from Selleckchem. Ro-106-9920 (#1178), TPCA-1 (#2559), PS1145 (#4569), Bay11-7082 (#1744), Ruxolinitib (#7048) and Stattic (#2798) were purchased from TOCRIS bioscience. TAK-242 (# 614316) was purchased from Millipore. Etanercept was supplied by Pfizer.

Clanchy F.I., Huang Y.S., Ogbechi J., Darlington L.G., Williams R.O, & Stone T.W. (2022). Induction of IDO1 and Kynurenine by Serine Proteases Subtilisin, Prostate Specific Antigen, CD26 and HtrA: A New Form of Immunosuppression?. Frontiers in Immunology, 13, 832989.

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RANKL-Induced Osteoclastogenesis Assay

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Recombinant mouse receptor activator of nuclear factor kappaβ ligand (RANKL) was purchased from BioVision Inc. (Milpitas, CA, US). Oleoyl-L-α-lysophosphatidic acid sodium salt (LPA) was from Sigma (US). LPA receptor-1/2/3inhibitor Ki16425 was bought from Santa Cruz Biotechnology (CA, US). 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) were purchased from Amresco (Albany, NY, US). PD98059, SB203580, SP600125, LY294002, BAY-11-7082, Go6976 and GF109203X were purchased from Tocris Bioscience (Bristol, UK), Y27632 from Cayman Chemical (Ann Arbor, MI, US). Phospho-(pan) PKC (Ser660) antibody was from Cell Signaling Technology (Boston, US). Anti-β-actin was obtained from Santa Cruz Biotechnology. Peroxidase-AffiniPure goat anti-rabbit IgG (H+L) and peroxidase-AffiniPure goat anti-mouse IgG (H+L) were supplied from Jackson ImmunoResearch (Baltimore, US).

Nam J.S., Sharma A.R., Nguyen L.T., Jagga S., Lee Y.H., Sharma G, & Lee S.S. (2018). Lysophosphatidic acid enhances breast cancer cells-mediated osteoclastogenesis. The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology, 22(5), 503-511.

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