Dextran Sulfate Sodium Salt (DSS) (Cat#9011-18-1) was purchased from MP Biomedicals (Irvine, CA). Specific antibodies including ZO-1 (1:1000; #AF5145; RRID:AB_2837631), Occludin (1:1000; #DF7504; RRID:AB_2841004), Claudin-3 (1:1000; #AF0129; #RRID:AB_2833313), p65 (1:1000; #BF8005:AB_2846809), p-p65 (1:1000; AB_2834435:AF2006), IKBα (1:1000; #AF5002:AB_2834792), p-IKBα (1:1000; #AB_2834433:AF2002), ERK (1:1000; #AB_2833336:AF0155), p-ERK (1:1000; #AB_2834432:AF1015), JNK (1:1000; #AF6318:AB_2835177), p-JNK (1:1000; #AF3318:AB_2834737), p38 (1:1000; #BF8015:Q16539), p-p38 (1:1000; #AF4001:AB_2835330) and GAPDH (1:1000; #AF7021; AB_2839421) were purchased from Affinity Biosciences (OH, USA). Tumor necrosis factor (TNF)-α (Cat# 430915), interleukin (IL)-1β (Cat# 432615), and interleukin (IL)-6 (Cat# 431307) enzyme-linked immunosorbent assay (ELISA) kits were obtained from Biolegend (San Diego, California, USA). Myeloperoxidas (MPO) (A044-1-1), Superoxide Dismutase (T-SOD) (A001-3-2), Malondialdehyde (MDA) (A003-1-2), Glutathione Peroxidase (GSH-PX) (A005-1-2), and Catalase (CAT) (A007-1-1) assay kit was bought from Nanjing Jiancheng Bioengineering Institute (Nanjing, China).
Tumor necrosis factor α
Manufactured by BioLegend
Sourced in United States
Tumor necrosis factor (TNF)-α is a cytokine that plays a central role in inflammation and immune response. It is involved in the regulation of a wide spectrum of biological processes, including cell proliferation, differentiation, apoptosis, lipid metabolism, and coagulation.
Automatically generated - may contain errors
Lab products found in correlation
Occludin, by Affinity Biosciences (2 mentions)
Claudin 3, by Affinity Biosciences (2 mentions)
Interleukin il 1β, by BioLegend (2 mentions)
P erk, by Affinity Biosciences (1 mentions)
P jnk, by Affinity Biosciences (1 mentions)
Dextran sulfate sodium salt dss, by MP Biomedicals (1 mentions)
P ikbα, by Affinity Biosciences (1 mentions)
Gapdh, by Affinity Biosciences (1 mentions)
Neomycin, by Merck Group (1 mentions)
Ampicillin, by Merck Group (1 mentions)
Vancomycin, by Merck Group (1 mentions)
Metronidazole, by Merck Group (1 mentions)
β actin, by Affinity Biosciences (1 mentions)
Cox 2, by Cell Signaling Technology (1 mentions)
P pkcδ, by Cell Signaling Technology (1 mentions)
P plcγ1, by Cell Signaling Technology (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Nf κb p65, by Cell Signaling Technology (1 mentions)
Chlorogenic acid, by Merck Group (1 mentions)
Dexamethasone (dex), by Merck Group (1 mentions)
6 protocols using tumor necrosis factor α
1
Inflammatory Bowel Disease Protocol
2
Inflammatory Bowel Disease Treatment Protocol
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Inulin was obtained from Yuanyebio Biotechnology Co., Ltd, (S11143, Shanghai, China). Ampicillin (Cat# A5354), neomycin (Cat# N6386), metronidazole (Cat# 16677), vancomycin (Cat# V2002) were bought from Sigma Aldrich (St. Louis, MO, USA). Specific antibodies including ZO-1 (1:1000; #AF5145; RRID: AB_2837631), Occludin (1:1000; #DF7504; RRID: AB_2841004), Claudin-3 (1:1000; #AF0129; RRID: AB_2833313) and β-actin (1:1000; #AF7018; RRID: AB_2839420) were purchased from Affinity Biosciences (OH, USA). S100A8 (#70802) and HDAC3 (#60538) antibody were bought from Cell Signaling Technology (Boston, USA). Acetylated-H3 antibody (Cat# ab47915; RRID: AB_873860) was purchased from Abcam (Cambridge, England). Tumor necrosis factor (TNF)-α (Cat# 430915) and interleukin (IL)-1β (Cat# 432615) enzyme-linked immunosorbent assay (ELISA) kits were obtained from Biolegend (San Diego, California, USA). Myeloperoxidase (MPO) (A044-1-1) assay kit was bought from Nanjing Jiancheng Bioengineering Institute (Nanjing, China,).
3
HPLC Analysis of Bioactive Compounds
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Standard samples (chlorogenic acid and γ-aminobutyric acid (GABA)) for high-performance liquid chromatography (HPLC), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) solution, lipopolysaccharide (LPS), vitamin C, and dexamethasone were obtained from Sigma-Aldrich (St. Louis, Missouri, USA). Recombinant human interferon (IFN)-γ and tumor necrosis factor (TNF)-α were obtained from BioLegend (San Diego, CA, USA). Fetal bovine serum (FBS), Dulbecco’s modified Eagle medium (DMEM), and Roswell Park Memorial Institute (RPMI) 1640 medium were purchased from Gibco BRL (Rockville, MD, USA). Enzyme-linked immunosorbent assay (ELISA) kit for histamine was obtained from Abcam (Cambridge, UK) for histamine. Antibodies against p-p38, p38, p-ERK, ERK, p-JNK, JNK, p-Lyn, Lyn, p-Syk, Syk, p-Fyn, Fyn, p-PLCγ1, PLCγ1, p-PKCδ, PKCδ, NF-κB p65, Lamin B, Nrf2, HO-1, NQO1, iNOS, COX-2, and β-actin were purchased from Cell Signaling Technology (Danvers, MN, USA) or Santa Cruz (Dallas, TX, USA).
4
Adenosine Receptor Modulation Protocol
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Adenosine receptor selective agents included the A1R agonist, N6-cyclopentyladenosine (CPA, Sigma-Aldrich, St. Louis, MO, USA); A1R antagonist; 8-cyclopentyl-1,3-dipropylxanthine (DPCPX, Funakoshi, Tokyo); A2AR agonist, CGS21680 (Cosmo Bio, Tokyo, Japan); and A2AR antagonist, ZM241385 (Funakoshi) [67 (link)]. The rat recombinant interferon γ (IFNγ) and tumor necrosis factor α (TNFα) were purchased from Biolegend (San Diego, CA, USA) and R&D Systems (Minneapolis, MN, USA), respectively. Carbamazepine (CBZ) was obtained from Tokyo Chemical Industry (Tokyo, Japan). The AMPA/glutamate receptor (AMPA-R) agonist, 2-amino-3-(3-hydroxy-5-methyl-isoxazol-4-yl) propanoic acid (AMPA), inositol trisphosphate receptor (IP3-R) agonist, adenophostin-A (AdA), and non-selective glutamate transporter inhibitor, trans-4-carboxy-L-proline (PDC) were obtained from Wako (Osaka, Japan), Cosmo Bio and Funakoshi, respectively.
AdA, AMPA, PDC, IFNγ, and TNFα were dissolved in cell culture medium directly. CGS21680, DPCPX, ZM241385, and CBZ were initially dissolved at 10 mM in dimethyl sulfoxide. The final concentrations of dimethyl sulfoxide were lower than 0.2% (vol/vol). CPA was initially dissolved at 10 mM in 1N HCl.
AdA, AMPA, PDC, IFNγ, and TNFα were dissolved in cell culture medium directly. CGS21680, DPCPX, ZM241385, and CBZ were initially dissolved at 10 mM in dimethyl sulfoxide. The final concentrations of dimethyl sulfoxide were lower than 0.2% (vol/vol). CPA was initially dissolved at 10 mM in 1N HCl.
5
T-cell Subset Characterization by Flow Cytometry
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T-cell subsets were described based on the surface expression of CD45RA and CD62L, dividing the cells into 4 subsets; naïve (CD45RA+CD62L+), terminally differentiated effector memory or TEMRA (CD45RA+CD62L−), central memory (CD45RA−CD62L+), and effector memory (CD45RA−CD62L−; supplemental Figure 1 , available on the Blood website).
Cells were stimulated with eBioscience cell stimulation cocktail (Thermo Fisher Scientific) and fixed and permeabilized using a staining buffer set (Thermo Fisher Scientific) for intracellular cytokine staining. Cells were stained with conjugated antibodies against granzyme B, interferon gamma (IFN-γ), interleukin 2 (BD Biosciences), and tumor necrosis factor α (TNFα) (BioLegend).
Stained cells were analyzed using a BD LSR II cytometer (BD Biosciences). BD FACSDIVA software (BD Biosciences) was used to acquire events on the cytometer, and FlowJo software (Tree Star) and FCS Express 6 Flow Cytometry Research Edition (De Novo Software) were used to analyze acquired data. In total, 32 patients were included in the immunophenotype analysis.
Cells were stimulated with eBioscience cell stimulation cocktail (Thermo Fisher Scientific) and fixed and permeabilized using a staining buffer set (Thermo Fisher Scientific) for intracellular cytokine staining. Cells were stained with conjugated antibodies against granzyme B, interferon gamma (IFN-γ), interleukin 2 (BD Biosciences), and tumor necrosis factor α (TNFα) (BioLegend).
Stained cells were analyzed using a BD LSR II cytometer (BD Biosciences). BD FACSDIVA software (BD Biosciences) was used to acquire events on the cytometer, and FlowJo software (Tree Star) and FCS Express 6 Flow Cytometry Research Edition (De Novo Software) were used to analyze acquired data. In total, 32 patients were included in the immunophenotype analysis.
6
Analyzing PSMP Signaling in Prostate Cancer
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Antibodies against extracellular signal-regulated kinase (ERK), phosphorylated ERK (p-ERK), AKT, p-AKT, E-cadherin, and β-actin were purchased from CST. The polyclonal antibody and neutralizing antibody against PSMP were produced in our lab as previously described (9 (link)). PC3, DU145, and THP-1 cell lines were obtained from the American Type Culture Collection (ATCC) and cultured following the recommended protocol. granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-4 (IL4), tumor necrosis factor-α (TNF-α), and lipopolysaccharide (LPS) were purchased from BioLegend. Enzyme-linked immunosorbent assay (ELISA) kits to detect IL1β, IL6, IL10, interferon-γ (IFN-γ), and CCL2 were from eBioscience. The PSMP detection method was previously described (10 (link)).
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